jess, Bart Haynes words are a little less conservative -


Method of inducing neutralizing antibodies to human immunodeficiency virus


Bart Haynes:

* "IS1 is an antibody that can be safely used as a therapeutic Mab for treatment of HIV infected subjects"


http://appft1.uspto.gov/netacgi/nph-Parser?Sect1=PTO2&Sect2=HITOFF&u=%2Fnetahtml%2FPTO%2Fsearch-adv.html&r=2&p=1&f=G&l=50&d=PG01&S1=%28%22haynes+barton%22.IN.%29&OS=in/%22haynes+barton%22&RS=IN/%22haynes+barton%22


.....................


Bart Haynes:

* "both IS1 and IS4 neutralized 7/7 primary isolates tested"


* "both IS1 and IS4 Mabs reacted strongly with the surface of virus infected T cells."




Supported by the Center for HIV/AIDS Vaccine Immunology AI067854,
CHAVI 005 protocol.

http://www.hivvaccineenterprise.org/_dwn/Late_Breaker_Abstracts.pdf


........................


Induction of Plasma (TRAIL), TNFR-2, Fas Ligand and Plasma Microparticles After
HIV-1 Transmission: Implications for HIV-1 Vaccine Design



Bart Haynes:

* "Significant plasma TRAIL elevations occurred a mean of 7.2 days before the
peak of plasma viral load (VL), while TNFR2, Fas ligand and microparticle elevations
occurred coincident with maximum VL. Microparticles have been previously shown to
mediate immunosuppressive effects on T cells and macrophages. We found that T cell
apoptotic microparticles also potently suppressed in vitro IgG and IgA antibody
production by memory B cells."


* "Microparticles (MPs) are small
membrane-bound vesicles that are released from the surface of apoptotic cells by
exocytic or budding processes; as such, MPs bear cell surface markers and can bind
annexin V because of the expression of phosphatidylserine"


* "MPs have immunomodulatory activities and can promote immune
cell death; exosomes are also immunologically active, can suppress immune responses
(20,34,42,55), and have been reported elevated in chronic HIV-1 "


* "suppression of immune responses can be mediated by T cell MPs (32,34,35).
CXCR4+ and CCR5+ MPs can transfer co-receptors to co-receptor negative cells, making
them susceptible to infection by HIV-1 (48,57). Phagocytosis of MPs by macrophages
releases TGF-beta, prostaglandin E2 and IL-10 that can inhibit antigen-specific T and B cell
responses (20,35,42). In this regard, Estes et al. have shown dramatic increases in lymph
node TGF-beta and IL-10 on day 12 following SIV infection (22). Importantly, we have
demonstrated that PBMC and tonsillar cell MPs can directly inhibit memory B cell activation"


* "both Fas ligand and TRAIL are incorporated into MPs (37,53). Fas ligand
expressing MPs can directly induce apoptosis in nearby cells (20,37,53), and activated T cells
can be the target of Fas ligand-mediated proapoptotic microvesicles "


* "it is likely that MPs are responsible for the observed B
cell suppressive activity seen in vitro in Figure 8. In the setting of HIV-1 infection where
both activation and apoptosis occur, however, MPs and exosomes may act concomitantly,
with exosomes suppressing immune responses (2,7,15,61), and MPs contributing to both
immune suppression and cell death"


http://jvi.asm.org/cgi/content/full/82/15/7700?view=long&pmid=18508902


----------


j

Other pertinent posts -


http://investorshub.advfn.com/boards/read_msg.aspx?message_id=29699542

http://investorshub.advfn.com/boards/read_msg.aspx?message_id=29699665

http://investorshub.advfn.com/boards/read_msg.aspx?message_id=29512347

http://investorshub.advfn.com/boards/read_msg.aspx?message_id=29516189

http://investorshub.advfn.com/boards/read_msg.aspx?message_id=29445060

lafont, Yes, here -

http://investorshub.advfn.com/boards/read_msg.aspx?message_id=29699506


j

New York Times TODAY ----------

http://www.nytimes.com/2008/07/18/health/18vaccine.html?_r=2&hp&oref=slogin&oref=slogin

snip-

In an unrelated development, researchers at Duke reported new findings showing that H.I.V. stuns the immune system earlier than scientists prevously understood. The window of opportunity in stopping H.I.V. may be a matter concerning the first few days — not weeks — after the virus enters the body, a team headed by Dr. Barton Haynes reported in The Journal of Virology. The findings were based on a study of 30 individuals newly infected with H.I.V. The study was paid for by the National Institutes of Health.


---------------


If you've been following my posts, you'll know that this is referring to Haynes work over the past couple years which PROVES THAT EXPOSED PS IS THE CULPRIT, work which has culminated so far with the following results -


What the NYT is specifically referring to above is the recent Gasper-Smith / Haynes paper, which shows that EXPOSED PS MUST BE DEALT WITH! The paper referred to today by the New York Times explains how a future successful vaccine MUST INDUCE ANTI-PS abs to counter the massive apoptosis and apoptotic debris (PS-exposing microparticles) that shut off immune responses.


His recent patent applications that I posted provide further details-



most recently- the US patent application I found and posted here two days ago talks about PEREGRINE'S ANTI-PS being very effective at neutralizing the virus. He then, (for the first time) discusses using PEREGRINE'S ANTI-PS AS AN HIV THERAPEUTIC.


In a nut-shell, Haynes (and Gates and CHAVI) are very focussed on exposed PS as the culprit they need to deal with.


---------



Here again are excerpts from both the WIPO patent app, as well as the US patent app. -


from Bart Haynes WIPO patent application -------------


.........Thus, HTV virions and HIV envelope can directly induce T cell death in AHI, soluble TRAIL can bind to uninfected cells and induce death in AHI, and with both HTV infection of cells and with massive apoptosis, high levels of phosphatidylserine containing cells and particles likely abound in AHI. It has recently been shown that PD-I (programmed death molecule- 1) is present on the surface of human B cells in chronic HIV infection. This suggests that human B cells are primed for apoptosis in HIV infection (Figure 6). HIV specific CD8+ T cell PD-I expression correlates with a CD8+ T cell response to poorly controlled chronic HIV infection (Petrovas et al, JEM 203: 2281 (2006)). Phosphatidylserine (PS) on the surface of HIV infected cells and virions has been found (Figure 7) and Callahan et al have found PS is a cofactor for HIV infection of monocytes (Callahan, J. Immunol 170:4840 (2003)). PS-dependent ingestion of apoptotic cells promotes TGF-beta secretion (Huynh et al, J. Clin. Invest. 109:41 (2002)) and interaction between PS and PS receptor inhibits antibody responses in

vivo (Hoffman et al, J. Immunol. 174:1393 (2005)) [ (jazzbeerman's "required reading #1") ] . INF-alpha, an anti-viral cytokine, sensitizes lymphocytes to apoptosis (Carrero et al, JEM 200:535 (2004)). There are increases in PS+ shed membrane particles in chronic HIV infection (Aupeix et al, J. Clin. Invest. 99:1546 (1997).........

Thus, the massive apoptosis that occurs with acute HIV infection with resulting release of TRAIL, mediation of apoptosis via FAS-FASL interactions, and release of PS containing viral and other particles all conspire to initially immunosuppress the host, preventing rapid protective B cell responses.

The present invention includes strategies to prevent apoptosis that include, but are not limited to, the use of PS-containing HIV immunogens, such as PS liposomes, either with or without CON-S or CON-T gpl40 or HTV env epitopes associated with the liposomes, such as 2F5-GTH1 peptide lipid conjugates (Figures 8A and 8B) administered with adjuvants to break tolerance and induce anti-PS antibodies that inhibit PS-CD36 interactions. Alternatively, recombinant CD36 can be targeted in order to raise anti-CD36 antibodies, preferably, both anti-PS or anti-CD36 antibodies are induced at mucosal sites to prevent apoptotic mediated immune suppression.


--------

US PATENT APP-



So, Haynes wants an HIV vaccine to induce anti-PS --------


WHY?.........

Obviously, because he's run passive mab experiments with anti-PS that are very promising. Experiments that show that anti-PS neutralizes HIV.


SURE ENOUGH --------------------


Now we know this to be a fact, as discussed in his recent US patent application, in which he discusses IS1 neutralizing HIV, (seven out of seven strains in one report).

For the first time, besides talking about the essential role/goal of anti-PS in a vaccine,
he's now talking about anti-PS as an HIV therapeutic-


'IS1 is an antibody that can be safely used as a therapeutic Mab for treatment of HIV infected subjects'

'can broadly neutralize HIV in an unprecedented manner'

'reactivity with the HIV envelope is not a prerequisite for neutralization in these antibodies'

'if HIV has evolved to escape the host immune response by making the immune system blind to it, other infectious agents may have evolved similarly. That is, this may represent a general mechanism of escape. That being the case, approaches comparable to those described herein can be expected to be useful in the treatment of such other agents well.'


----------


I am looking forward to reading all about it.....

IT'S HAPPENING :)


j

Al,

IMO, so early in development, as for up-front money, not much.


j

So, Haynes wants an HIV vaccine to induce anti-PS --------


WHY?.........

Obviously, because he's run passive mab experiments with anti-PS that are very promising. Experiments that show that anti-PS neutralizes HIV.


SURE ENOUGH --------------------


Now we know this to be a fact, as discussed in his recent US patent application, in which he discusses IS1 neutralizing HIV, (seven out of seven strains in one report).

For the first time, besides talking about the essential role/goal of anti-PS in a vaccine,
he's now talking about anti-PS as an HIV therapeutic-


'IS1 is an antibody that can be safely used as a therapeutic Mab for treatment of HIV infected subjects'

'can broadly neutralize HIV in an unprecedented manner'

'reactivity with the HIV envelope is not a prerequisite for neutralization in these antibodies'

'if HIV has evolved to escape the host immune response by making the immune system blind to it, other infectious agents may have evolved similarly. That is, this may represent a general mechanism of escape. That being the case, approaches comparable to those described herein can be expected to be useful in the treatment of such other agents well.'


----------


I am looking forward to reading all about it.....

IT'S HAPPENING :)


j

from Bart Haynes WIPO patent application -------------


.........Thus, HTV virions and HIV envelope can directly induce T cell death in AHI, soluble TRAIL can bind to uninfected cells and induce death in AHI, and with both HTV infection of cells and with massive apoptosis, high levels of phosphatidylserine containing cells and particles likely abound in AHI. It has recently been shown that PD-I (programmed death molecule- 1) is present on the surface of human B cells in chronic HIV infection. This suggests that human B cells are primed for apoptosis in HIV infection (Figure 6). HIV specific CD8+ T cell PD-I expression correlates with a CD8+ T cell response to poorly controlled chronic HIV infection (Petrovas et al, JEM 203: 2281 (2006)). Phosphatidylserine (PS) on the surface of HIV infected cells and virions has been found (Figure 7) and Callahan et al have found PS is a cofactor for HIV infection of monocytes (Callahan, J. Immunol 170:4840 (2003)). PS-dependent ingestion of apoptotic cells promotes TGF-beta secretion (Huynh et al, J. Clin. Invest. 109:41 (2002)) and interaction between PS and PS receptor inhibits antibody responses in

vivo (Hoffman et al, J. Immunol. 174:1393 (2005)) [ (jazzbeerman's "required reading #1") ] . INF-alpha, an anti-viral cytokine, sensitizes lymphocytes to apoptosis (Carrero et al, JEM 200:535 (2004)). There are increases in PS+ shed membrane particles in chronic HIV infection (Aupeix et al, J. Clin. Invest. 99:1546 (1997).........

Thus, the massive apoptosis that occurs with acute HIV infection with resulting release of TRAIL, mediation of apoptosis via FAS-FASL interactions, and release of PS containing viral and other particles all conspire to initially immunosuppress the host, preventing rapid protective B cell responses.

The present invention includes strategies to prevent apoptosis that include, but are not limited to, the use of PS-containing HIV immunogens, such as PS liposomes, either with or without CON-S or CON-T gpl40 or HTV env epitopes associated with the liposomes, such as 2F5-GTH1 peptide lipid conjugates (Figures 8A and 8B) administered with adjuvants to break tolerance and induce anti-PS antibodies that inhibit PS-CD36 interactions. Alternatively, recombinant CD36 can be targeted in order to raise anti-CD36 antibodies, preferably, both anti-PS or anti-CD36 antibodies are induced at mucosal sites to prevent apoptotic mediated immune suppression.



---------


j

free,

Yes. That paper http://www.sciencemag.org/cgi/content/abstract/sci;308/5730/1906?ck=nck made a big splash. Haynes presented his findings at CROI 2006. It was the work that resulted in Haynes getting the director job at CHAVI, and also having the big pile of Gates money thrown at him.

Since then, we now see that he has moved from thinking that successful broadly-neutralizing mabs need to be polyspecific, (binding both a viral epitope as well as a phospholipid), to saying that successful anti-HIV therapeutic antibodies need only bind the lipid! (As discussed in the patent application I posted the other day which is all about PEREGRINE'S Pojen Chen's mabs isolated from an autoimmune patient back in 1999).

[0105] ....That IS1 neutralized HIV evidences the facts that: a) humans can make non-pathogenic anti-lipid antibodies that neutralize HIV, and b) IS1 is an antibody that can be safely used as a therapeutic Mab for treatment of HIV infected subjects or in the setting of post-exposure prophylaxis of subjects following needle, sexual or other exposure to HIV or HIV infected materials.


[0108] Alving and colleagues have made a mouse mab against phosphatidyl inositol phosphate and have shown that it neutralizes HIV in a PBMC assay. What the present studies show is that humans can spontaneously make anti-lipid antibodies and that these antibodies can broadly neutralize HIV in an unprecedented manner.



[0109] Summarizing, autoimmune disease patients can make antibodies that bind to virus-infected cells and, presumably, to budding HIV virions by virtue of their reactivity to HIV membranes and host membranes. Certain anti-lipid antibodies from autoimmune disease patients can also react with the Envelope trimer (such as IS6) but not all of the antibodies react also with the trimer (i.e., IS1 and IS4 do not react). Therefore, reactivity with the HIV envelope is not a prerequisite for neutralization in these antibodies.



You may remember a year or two ago, discussing Haynes work at the time looking into polyspecific abs, I wrote a post titled,
"It's the Lipid Stupid".

Well, it seems he's now on board :)

j

-------------------------
http://investorshub.advfn.com/boards/read_msg.aspx?message_id=18077846

Posted by: jazzbeerman Date: Wednesday, March 21, 2007 12:20:19 PM
In reply to: None Post # of 26178

Haynes / Alving..... It's the LIPID stupid.


I still wait for Haynes to say something similar to Alving at Walter Reed, basically, that perhaps polyspecificity is not necessary... (and that it's not the viral epitope, but the PS-binding region that's the more important neutralizing aspect of the 4E10 mab)...
Haynes has discovered (over the past two or three years) that the few rare broadly-neutralizing HIV mabs isolated from a few lucky infected folks, besides binding HIV epitopes, also bind anionic phospholipids. Haynes has been putting forth the idea that binding anionic phospholipids may be a necessary characteristic of broadly-neutralizing HIV abs, which obviously makes a lot of sense, (for broadly-netralizing HIV, and any other virus that wears anionic lipids on the outside, which is every enveloped virus)..

ALVING (at Walter Reed, head of HIV vaccine development), on the other hand, has ALSO looked into the effect of abs against the phospholipids WITHOUT also binding viral epitopes... They worked. Alving's ab to PI phosphate neutralized HIV, just like 4E10, but Alving's ab was strictly binding the lipid, not any viral epitope.

Apparently Haynes has seen similar results? (with bavituximab), but simply hasn't talked about it or published anyting about it, yet...?

Steve King, PPHM CEO, from the latest conference call, with regard to the soon to begin bavituximab HIV clinical trial:

"...Our interest in this patient population [HVC/HIV] has been further stimulated by solid evidence, through our collaborators at Duke, that Bavituximab binds to HIV virus, binds to HIV-infected cells, and may have potent neutralizing effects on the virus."


It's important to note here that others have recently found that simply blocking PS with annexin V strongly inhibits HIV replication.

Gaipl theorizes about the proven immunity to specific tumors noted in their experiments with annexin V, saying that by covering PS, the apoptotic tumor cells are not phagocytosed, and hence are allowed to further fall apart, leaking more danger signals. This prolonged death process, this delayed clean-up, ignored by macrophages long enough for the cell's nasty contents to start slipping out, is enough to initiate an inflammatory response, firing up the DC's etc., resulting in the innate/adaptive response.

It makes me wonder, while that's a fascinating and promising way to stimulate the immune system to fight cancer and viral infections, if it isn't also increasing the liklihood/possibility of autoantibodies / antiphospholipid syndrome, as lupus and other diseases have been found to be related to delayed clearance of apoptotic cells..



It seems likely that covering PS with an antibody could show similar results to the now proven mechanism about to be discussed by UTSW scientists at AACR, in which bavi-opsonized apopototic tumor cells were efficiently phagocytosed by DCs through FcgR on DC's (causing a tumor-specific T cell immune response).


j

Malaria Exploits Exposed PS ------




Cytoadherence of Malaria-Infected Red Blood
Cells Involves Exposure of Phosphatidylserine

http://content.karger.com/ProdukteDB/produkte.asp?Aktion=ShowPDF&ProduktNr=224332&Ausgabe=228865&ArtikelNr=67908&filename=67908.pdf


Introduction

Malaria, one of the most life-threatening infectious
diseases in the world, causes 1.5-2.7 million deaths
annually [1]. Of the four species of human malaria
parasites, Plasmodium falciparum causes the severest of
symptoms and the greatest number of deaths. Adherence
of red cells infected with the mature stages of the parasite
to endothelial cells (EC) lining the postcapillary venules,
especially in the deep tissues, enables the parasites to
avoid splenic clearance and parasite growth is favored
by an hypoxic microenvironment. The adherence of
P. falciparum-infected red cells also results in the
occlusion of microvessels and can lead to
unconsciousness, coma, and in some cases, death. The
virulent nature of P. falciparum has been attributed to
mechanical obstruction by adherent infected red cells [2].
In this context, an understanding of the mechanisms of
the adherence of malaria-infected red cells may lead to
the development of anti-adhesive reagents and new
therapies.

......

The asymmetry of phospholipids in the lipid bilayer
of the plasma membrane is maintained by several membrane
enzymes such as scramblase, translocase and
flippase [28]. Disruption of phospholipid asymmetry
results in the exposure of phosphatidylserine (PS)
molecules, which are normally located in the inner leaflet
of the phospholipid bilayer. We and other investigators
have shown that PS is exposed on the malaria-infected
red cell surface coincident with parasite development [29-
31]. PS liposomes or PS-exposed cells bind to CD36 [32]
and TSP [33] both of which serve as receptors for the
adherence of P. falciparum-infected red cells [34, 35].
Here, we demonstrate that PS exposure on the exofacial
surface of the P. falciparum -infected cell is involved in
the binding to CD36 and TSP.

.......

Discussion
Drastic changes in the red blood cell membrane are
induced during the intracellular maturation of the malarial
parasite, P. falciparum. These include visible changes in
the shape of the red cell, the appearance of electron-dense
protrusions (called knobs) [3, 49], increased membrane
permeability [50, 51], decreased deformability [52],
insertion of parasite-derived proteins [8-10], changes in
the composition and oxidative damage of membrane
lipids [53, 54], a reduction in cholesterol content [30],
as well as degradation of membrane proteins [55].

........

Further,
Facer et al. [63] detected elevated levels of antiphospholipid
antibodies in the serum of P. falciparuminfected
patients using an ELISA method, and found the
highest IgG and IgM binding was to PS and other anionic
phospholipids, indicating that infected red cells expose
PS on their surface in vivo. In that report, the percentage
of anti-PS IgG or IgM positive serum of P. falciparuminfected
patients as 89% and 79%, respectively. Thus,
the accumulating data support PS exposure in P.
falciparum infected red cells.

.......

The precise mechanism of PS exposure on
P.falciparum-infected red cells remains undefined.
Several investigators have demonstrated that treatment
of red cells with various oxidants, such as
phenylhydrazine [64], hydrogen peroxide [65], or
diamide [66], also results in PS exposure. And, PS
exposure has been demonstrated in red cells from patients
with sickle cell anemia [36, 67, 68], thalassemia [69],
diabetes [70], all of which may be under oxidative stress
[71-73]. The erythrocytic stages of Plasmodium as they
mature, digest hemoglobin, and release highly oxidative
iron-containing products. And, oxidized membrane lipids
have been detected in trophozoite- or schizont-infected
red cell membranes [54], coincident with the time we
and other investigators have detected PS exposure [29-
31]. It seems plausible that oxidative stress could act as
a trigger for PS exposure on the surface of the malariainfected
red cell.

.......

Since PS exposure was detected in the various strains of
falciparum malaria we tested, as well as strains used by
other workers, we contend that PS exposure can
contribute to the universal binding of infected cells to
CD36 and TSP. Our findings (i.e., PS exposure and PSmediated
infected-cell binding to TSP) is also compatible
with the suggestion that PS is involved in abnormal sickle
red cell binding to TSP [76, 77].

.......

Further investigation of the structural requirement of
inhibition of PS-mediated infected-cell binding, as well
as a determination of the relative contributions of PfEMP-
1, band 3 and PS to cytoadherence, may assist in the
development of novel and effective anti-adhesive agents.
In conclusion, PS exposure, modification in band 3
protein, and PfEMP-1 all contribute to cytoadherence/
sequestration and disease pathogenesis in P. falciparum
malaria.




----------




Evolution has favored pathogenesis that resembles apoptosis,

j







High levels of anti-phospholipid antibodies in uncomplicated and severe Plasmodium falciparum and in P. vivax malaria.

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=8306506&dopt=Abstract

Facer CA, Agiostratidou G.

London Hospital Medical College, UK.


High levels of anti-phospholipid antibodies in uncomplicated and severe Plasmodium falciparum and in P. vivax malaria.



The majority (75%) of adult patients with uncomplicated Plasmodium falciparum and P. vivax malaria are positive for anti-phospholipid antibodies (aPLA) as demonstrated by ELISA using a panel of anionic and cationic phospholipids. The highest IgG and IgM binding was to the anionic phospholipids, phosphatidylserine (PS), phosphatidic acid (PA) and cardiolipin (CL), but excluding phosphatidylinositol (PI) to which only low antibody levels were found. Comparison of the mean IgG and IgM aPLA showed a trend for anti-PA > CL > PS > PC > PE > PI. Anti-PI levels were compared in two groups of African children, one group with non-severe and the other with severe (cerebral) falciparum malaria. Children with cerebral disease had significantly lower IgM anti-PI. The results are discussed with the view that serum-derived aPLA may have a role in 'anti-disease' immune responses. Their possible role in the opsonization and phagocytosis of parasitized erythrocytes and in thrombocytopenia is also considered.




-------------------------






Upregulation of TGF-beta, FOXP3, and CD4+CD25+ regulatory T cells correlates with more rapid parasite growth in human malaria infection.


Immunity. 2005 Sep

Comment in:
Immunity. 2005 Sep;23(3):241-2.


Center for Clinical Vaccinology and Tropical Medicine, Nuffield Department of Clinical Medicine, Oxford University, Churchill Hospital, Oxford OX3 7LJ, United Kingdom.

Understanding the regulation of immune responses is central for control of autoimmune and infectious disease. In murine models of autoimmunity and chronic inflammatory disease, potent regulatory T lymphocytes have recently been characterized. Despite an explosion of interest in these cells, their relevance to human disease has been uncertain. In a longitudinal study of malaria sporozoite infection via the natural route, we provide evidence that regulatory T cells have modifying effects on blood-stage infection in vivo in humans. Cells with the characteristics of regulatory T cells are rapidly induced following blood-stage infection and are associated with a burst of TGF-beta production, decreased proinflammatory cytokine production, and decreased antigen-specific immune responses. Both the production of TGF-beta and the presence of CD4+CD25+FOXP3+ regulatory T cells are associated with higher rates of parasite growth in vivo. P. falciparum-mediated induction of regulatory T cells may represent a parasite-specific virulence factor.




------------




j

Trypanosoma Exploits Exposed PS ------


FEMS Microbiol Lett. 2006 Nov 1



Trypanosoma cruzi exposes phosphatidylserine as an evasion mechanism.

Damatta RA, Seabra SH, Deolindo P, Arnholdt AC, Manhaes L, Goldenberg S, de Souza W.

Laboratorio de Biologia Celular e Tecidual, Universidad Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil.

Chagas disease is caused by Trypanosoma cruzi and affects 18 million people in Central and South America. Here we analyzed the exposure of phosphatidylserine by the different forms of the parasite life cycle. Only the infective trypomastigotes, but not the epimastigotes or intracellular amastigotes, expose this phospholipid. This triggers a transforming growth factor beta signaling pathway, based on phosphorylated Smad 2 nuclear translocation, leading to iNOS disappearance in infected macrophages This macrophage deactivation favors the survival of this intracellular parasite. Thus, phosphatidylserine exposure may be used by T. cruzi to evade innate immunity and be a common feature of obligate intracellular parasites that have to deal with activated macrophages.


FULL PAPER:
http://www.blackwell-synergy.com/doi/full/10.1111/j.1574-6968.2006.00495.x?cookieSet=1


-----------



j

Schistosomes Exploit Exposed PS ------




Parasite immunity: Why worms are a turn off

Nature Reviews Immunology
http://www.signaling-gateway.org/update/updates/200301/nri989.html



Chistosome lyso-phosphatidylserine seems to signal through TLR2 to direct the induction of IL-10-secreting helper T cells, possibly accounting for the skewing to IL-10 production and T-cell hyporesposiveness that is seen during chronic schistosomiasis.


Researchers at Leiden University have identified a schistosome lipid that seems to interact with the innate immune system through a Toll-like receptor (TLR). Interestingly, rather than promoting T-cell activation, this interaction induces the development of T cells with immunosuppressive potential.


Schistosomes are parasitic worms that chronically infect more than 200 million people in developing countries. Owing to complex interactions between the host immune system and the parasite — the product of a long period of co-evolution — the schistosomes can survive for many years.

In recent years, lipids have emerged as important modulators of the innate immune system. So, van der Kleij and colleagues set out to investigate the effects of schistosome lipids on the host immune response. Lipids from Schistosoma mansoni were fractionated using an ion-exchange column and then tested for their capacity to induce cytokine production.

A fraction containing phosphatidylserine was found to affect the maturation of dendritic cells (DCs) such that they induced the development of T cells secreting the immunosuppressive cytokine interleukin-10 (IL-10) in vitro. In co-culture, these T cells suppressed the proliferation of autologous T cells, an effect that was blocked by IL-10-specific antibodies. This effect seems to be specific for phosphatidylserine, as other schistosome extracts failed to induce the production of IL-10 by T cells in vitro. Mammalian phosphatidylserine had no effect on DC maturation.

Members of the TLR family have been shown to recognize other pathogen-associated lipids, and so they were candidates receptors for interaction with the schistosome phosphatidyl fraction. Antibodies specific for TLR2 blocked the induction of IL-10-secreting T cells by phosphatidylserine in vitro, indicating that TLR2 might be involved in the recognition of schistosome phosphatidylserine. Phospholipase digestion of the phosphatidylserine lipid fraction reduced its TLR2-stimulating capacity, confirming that the active moiety is a phosphatidylserine and not a contaminant. Further fractionation of the worm phosphatidylserine by HPLC pinpointed the TLR2-activating and IL-10-promoting activity to a lyso-phosphatidylserine fraction.

So, schistosome lyso-phosphatidylserine seems to signal through TLR2 to direct the induction of IL-10-secreting helper T cells, which might account for the skewing to IL-10 production and T-cell hyporesposiveness that is seen during chronic schistosomiasis.





-----------------------




j

Toxoplasma Gondii Exploits Exposed PS -------






Biochem Biophys Res Commun. 2004 Nov 12


Toxoplasma gondii exposes phosphatidylserine inducing a TGF-beta1 autocrine effect orchestrating macrophage evasion.

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_...

Seabra SH, de Souza W, Damatta RA.

Laboratorio de Biologia Celular e Tecidual, Centro de Biociencias e Biotecnologia, Universidade Estadual do Norte Fluminense, 28013-600 Campos dos Goytacazes, RJ, Brazil.


Toxoplasmosis is a worldwide disease caused by Toxoplasma gondii. Activated macrophages control T. gondii growth by nitric oxide (NO) production. However, T. gondii active invasion inhibits NO production, allowing parasite persistence. Here we show that the mechanism used by T. gondii to inhibit NO production persisting in activated macrophages depends on phosphatidylserine (PS) exposure. Masking PS with annexin-V on parasites or activated macrophages abolished NO production inhibition and parasite persistence. NO production inhibition depended on a transforming growth factor-beta1 (TGF-beta1) autocrine effect confirmed by the expression of Smad 2 and 3 in infected macrophages. TGF-beta1 led to inducible nitric oxide synthase (iNOS) degradation, actin filament (F-actin) depolymerization, and lack of nuclear factor-kappaB (NF-kappaB) in the nucleus. All these features were reverted by TGF-beta1 neutralizing antibody treatment. Thus, T. gondii mimics the evasion mechanism used by Leishmania amazonensis and also the anti-inflammatory response evoked by apoptotic cells.




----------------------------




Toxoplasmosis-

http://www.cdc.gov/ncidod/dpd/parasites/toxoplasmosis/factsht_toxoplasmosis.htm




------------------------------



Evolution has favored pathogenesis that resembles apoptosis,

j

Leishmania Exploits Exposed PS ------



parasitic infection through "apoptotic mimicry"....






The Journal of Immunology, 2006, 176: 1834-1839
.


Mimicry of Apoptotic Cells by Exposing Phosphatidylserine Participates in the Establishment of Amastigotes of Leishmania (L) amazonensis in Mammalian Hosts

http://www.jimmunol.org/cgi/content/abstract/176/3/1834


João L. M. Wanderley*,{dagger}, Maria E. C. Moreira*, Aline Benjamin*,{dagger}, Adriana C. Bonomo*,{ddagger} and Marcello A. Barcinski2,*,§


Signaling through exposed phosphatidylserine (PS) is fundamental for the TGFbeta1-dependent, noninflammatory phagocytosis of apoptotic cells. This same mechanism operates in the internalization of amastigotes of Leishmania (L) amazonensis (L(L)a) in a process quoted as apoptotic mimicry. Now we show that the host modulates PS exposure by the amastigotes and, as a consequence, BALB/c mice-derived amastigotes expose significantly more PS than those derived from C57BL/6 mice. Due to this difference in the density of surface PS molecules, the former are significantly more infective than the latter, both in vivo, in F1 (BALB/c x C57BL/6) mice, and in vitro, in thioglycollate-derived macrophages from this same mouse strain. PS exposure increases with progression of the lesion and reaches its maximum value in amastigotes obtained at the time point when the lesion in C57BL/6 mice begins to decrease in size and the lesions in BALB/c mice are still growing in size. Synthesis of active TGFbeta1, induction of IL-10 message, and inhibition of NO synthesis correlate with the amount of surface PS displayed by viable (propidium iodide-negative) infective amastigote. Furthermore, we also show that, similar to what happens with apoptotic cells, amastigotes of L(L)a are internalized by macropinocytosis. This mechanism of internalization is consistent with the large phagolysosomes characteristic of L(L)a infection. The intensity of macrophage macropinocytic activity is dependent on the amount of surface PS displayed by the infecting amastigote.




------------------------


more Leishmania & PS



Braz J Med Biol Res. 2005 Jun;38(6):807-12. Epub 2005 Jun 1.

Apoptotic mimicry: an altruistic behavior in host/Leishmania interplay.


Wanderley JL, Benjamin A, Real F, Bonomo A, Moreira ME, Barcinski MA.

Instituto Nacional de Cancer, 20231-050 Rio de Janeiro, RJ, Brasil.



Apoptosis is the most common phenotype observed when cells die through programmed cell death. The morphologic and biochemical changes that characterize apoptotic cells depend on the activation of a diverse set of genes. Apoptosis is essential for multicellular organisms since their development and homeostasis are dependent on extensive cell renewal. In fact, there is strong evidence for the correlation between the emergence of multicellular organisms and apoptosis during evolution. On the other hand, no obvious advantages can be envisaged for unicellular organisms to carry the complex machinery required for programmed cell death. However, accumulating evidence shows that free-living and parasitic protozoa as well as yeasts display apoptotic markers. This phenomenon has been related to altruistic behavior, when a subpopulation of protozoa or yeasts dies by apoptosis, with clear benefits for the entire population. Recently, phosphatidylserine (PS) exposure and its recognition by a specific receptor (PSR) were implicated in the infectivity of amastigote forms of Leishmania, an obligatory vertebrate intramacrophagic parasite, showing for the first time that unicellular organisms use apoptotic features for the establishment and/or maintenance of infection. Here we focus on PS exposure in the outer leaflet of the plasma membrane--an early hallmark of apoptosis--and how it modulates the inflammatory activity of phagocytic cells. We also discuss the possible mechanisms by which PS exposure can define Leishmania survival inside host cells and the evolutionary implications of apoptosis at the unicellular level.





--------------------------------







Curr Biol. 2001 Nov 27;11(23):1870-3.

Apoptotic mimicry by an obligate intracellular parasite downregulates macrophage microbicidal activity.



de Freitas Balanco JM, Moreira ME, Bonomo A, Bozza PT, Amarante-Mendes G, Pirmez C, Barcinski MA.

Instituto de Ciencias Biomedicas, Universidade de Sao Paulo, Sao Paulo 05508-900, Brazil.

Programmed cell death by apoptosis of unnecessary or potentially harmful cells is clearly beneficial to multicellular organisms. Proper functioning of such a program demands that the removal of dying cells proceed without an inflammatory reaction. Phosphatidylserine (PS) is one of the ligands displayed by apoptotic cells that participates in their noninflammatory removal when recognized by neighboring phagocytes. PS ligation induces the release of transforming growth factor-beta (TGF-beta), an antiinflammatory cytokine that mediates the suppression of macrophage-mediated inflammation. In Hydra vulgaris, an organism that stands at the base of metazoan evolution, the selective advantage provided by apoptosis lies in the fact that Hydra can survive recycling apoptotic cells by phagocytosis. In unicellular organisms, it has been proposed that altruistic death benefits clonal populations of yeasts and trypanosomatids. Now we show that advantageous features of the apoptotic process can operate without death as the necessary outcome. Leishmania spp are able to evade the killing activity of phagocytes and establish themselves as obligate intracellular parasites. Amastigotes, responsible for disease propagation, similar to apoptotic cells, inhibit macrophage activity by exposing PS. Exposed PS participates in amastigote internalization. Recognition of this moiety by macrophages induces TGF-beta secretion and IL-10 synthesis, inhibits NO production, and increases susceptibility to intracellular leishmanial growth.





-----------------------------




Leishmaniasis

http://www.sbri.org/diseases/leishmaniasis.asp


Impact

Leishmaniasis is a parasitic disease transmitted by the bite of a sandfly that is infected with Leishmania parasites. Currently 350 million people in 88 countries around the world are threatened, and 12 million people are affected by leishmaniasis. Of the 1.5 – 2 million new cases of leishmaniasis estimated to occur annually, most occur in the tropics and subtropics, including the Middle East. In 2002, leishmaniasis reached epidemic levels in Afghanistan, with the World Health Organization calling for more funding and research for the disease. Leishmania/HIV co-infection is emerging as a serious new disease and it is increasingly frequent. It is considered a threat in southwestern Europe, such as Spain, Italy, France, and Portugal.


Symptoms

With the bite of an infected sandfly, Leishmania parasites are passed from one infected animal or human to others. Leishmaniasis is a spectrum of diseases, each distinctly manifested and all with potentially devastating consequences – disfigurement, damage to internal organs, death. Depending on the species of the infecting parasite, the spleen, liver, bone marrow, mucous membranes, and/or skin may be attacked. Leishmania donovani, the most dangerous of these, causes Kala azar, or visceral leishmaniasis, characterized by fever, severe weight loss and anemia. If left untreated, visceral leishmaniasis can lead to death.




--------------------------





j

re: "Duke" paper(s) --------------------


If you're wondering what type of stuff is going to be in the papers, look around this patent application for starters....




Obviously, Haynes and team will be detailing experiments that got Haynes to say crazy things like:



* Bart Haynes: "IS1 is an antibody that can be safely used as a therapeutic Mab for treatment of HIV infected subjects"
HUH?! REALLY?! HOW?! WHY ARE YOU SAYING THAT?!




* Bart Haynes: "these antibodies can broadly neutralize HIV in an unprecedented manner"
THEY DO?! HOW?! WHY IS IT UNPRECEDENTED?!
(answer: because nobody has previously aimed at host-cell phospholipids as deliberate targets for immunotherapy because it blatantly goes against the prevailing immunological paradigm, and it was only discovered very recently that PS is exposed on HIV, and that PS is essential to enveloped virus entry into cells).


* Bart Haynes: "reactivity with the HIV envelope is not a prerequisite for neutralization in these antibodies."
(see above comment).


* Bart Haynes: "if HIV has evolved to escape the host immune response by making the immune system blind to it, other infectious agents may have evolved similarly. That is, this may represent a general mechanism of escape. That being the case, approaches comparable to those described herein can be expected to be useful in the treatment of such other agents well."
HUH?! REALLY? OTHER INFECTIOUS AGENTS MAY SIMILARLY TRICK OUR IMMUNE SYSTEM?! LIKE WHAT?! OTHER ENVELOPED PS-EXPOSING VIRUSES?! MALARIA?! TUBERCULOSIS?! TRYPANOSOMA?! LEISHMANIA?! TOXOPLASMA GONDII?! SCHISTOSOMES?! AND THE SAME TYPE OF THERAPY LOOKS PROMISING THERE?! REALLY?! WHY DO YOU SAY THAT?!........





Evolution has favored pathogenesis that resembles apoptosis.


j

HIV, anti-PS, & the new Bart Haynes patent application --------------




1. A method of treating HIV comprising administering to a patient in need thereof an antibody derivable from a normal subject or from an autoimmune disease subject that binds to a lipid on the surface of HIV or on the surface of HIV-infected cells and thereby neutralizes HIV-1, wherein said antibody is administered in an amount sufficient to effect said treatment.

2. The method according to claim 1 wherein said antibody is derivable from an anti-phospholipid syndrome subject.

3. The method according to claim 1 wherein said antibody is non-pathogenic.

4. The method according to claim 1 wherein said antibody is IS1, IS4 or IS6, or binding fragment thereof.

5. The method according to claim 1 wherein said antibody is IS1, or binding fragment thereof.


[0105] ....That IS1 neutralized HIV evidences the facts that: a) humans can make non-pathogenic anti-lipid antibodies that neutralize HIV, and b) IS1 is an antibody that can be safely used as a therapeutic Mab for treatment of HIV infected subjects or in the setting of post-exposure prophylaxis of subjects following needle, sexual or other exposure to HIV or HIV infected materials.


[0108] Alving and colleagues have made a mouse mab against phosphatidyl inositol phosphate and have shown that it neutralizes HIV in a PBMC assay. What the present studies show is that humans can spontaneously make anti-lipid antibodies and that these antibodies can broadly neutralize HIV in an unprecedented manner.



[0109] Summarizing, autoimmune disease patients can make antibodies that bind to virus-infected cells and, presumably, to budding HIV virions by virtue of their reactivity to HIV membranes and host membranes. Certain anti-lipid antibodies from autoimmune disease patients can also react with the Envelope trimer (such as IS6) but not all of the antibodies react also with the trimer (i.e., IS1 and IS4 do not react). Therefore, reactivity with the HIV envelope is not a prerequisite for neutralization in these antibodies.


AND MY FAVORITE "broad implication" point mentioned by Haynes -

[0051] It will be appreciated from a reading of the foregoing that if HIV has evolved to escape the host immune response by making the immune system blind to it, other infectious agents may have evolved similarly. That is, this may represent a general mechanism of escape. That being the case, approaches comparable to those described herein can be expected to be useful in the treatment of such other agents well.


http://appft1.uspto.gov/netacgi/nph-Parser?Sect1=PTO2&Sect2=HITOFF&u=%2Fnetahtml%2FPTO%2Fsearch-adv.html&r=2&p=1&f=G&l=50&d=PG01&S1=%28%22haynes+Barton%22.IN.%29&OS=in/%22haynes+Barton%22&RS=IN/%22haynes+Barton%22


--------


j

jake,

the patent application is discussing the anti-PS mabs IS1, IS4,& IS6.

IS1, IS4, & IS6 are Pojen Chen's / University of California.

Chen showed up on PPHM's SRB listed under CORE TECHNOLOGIES last summer.

We just learned yesterday in the 10K, that last summer, PPHM licensed anti-PS mabs from University of California.
Then Chen showed up on PPHM's SRB.
Then the mabs started showing up in Duke's CHAVI/GATES HIV work.


Also, if you're interested, humanized Bavituximab is also named in that Haynes patent application.



j

8. *** AND MY FAVORITE SNIP!!!!!!!!!!! -



BART HAYNES US PATENT APPLICATION pt. 4



[0051] It will be appreciated from a reading of the foregoing that if HIV has evolved to escape the host immune response by making the immune system blind to it, other infectious agents may have evolved similarly. That is, this may represent a general mechanism of escape. That being the case, approaches comparable to those described herein can be expected to be useful in the treatment of such other agents well.


Bart Haynes:
" if HIV has evolved to escape the host immune response
by making the immune system blind to it,
other infectious agents may have evolved similarly. "

"That is, this may represent a general mechanism of escape."



Bart Haynes:
" if HIV has evolved to escape the host immune response
by making the immune system blind to it,
other infectious agents may have evolved similarly. "

"That is, this may represent a general mechanism of escape."



Bart Haynes:
" if HIV has evolved to escape the host immune response
by making the immune system blind to it,
other infectious agents may have evolved similarly. "

"That is, this may represent a general mechanism of escape."


------------

!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!

IT'S HAPPENING :)


j

7. *** BART HAYNES US PATENT APPLICATION pt. 2 -------------------


[0108] Alving and colleagues have made a mouse mab against phosphatidyl inositol phosphate and have shown that it neutralizes HIV in a PBMC assay. What the present studies show is that humans can spontaneously make anti-lipid antibodies and that these antibodies can broadly neutralize HIV in an unprecedented manner.


again -


Bart Haynes:
"these antibodies can broadly neutralize HIV in an unprecedented manner."


Bart Haynes:
"these antibodies can broadly neutralize HIV in an unprecedented manner."


Bart Haynes:
"these antibodies can broadly neutralize HIV in an unprecedented manner."



------------


It's happening :)

j

6. *** IS1 mab in new BART HAYNES US PATENT APPLICATION ---------------



An exciting patent application from Bart Haynes has recently hit the govt database -


http://appft1.uspto.gov/netacgi/nph-Parser?Sect1=PTO2&Sect2=HITOFF&u=%2Fnetahtml%2FPTO%2Fsearch-adv.html&r=2&p=1&f=G&l=50&d=PG01&S1=%28%22haynes+Barton%22.IN.%29&OS=in/%22haynes+Barton%22&RS=IN/%22haynes+Barton%22


Here are a few snips- get this.....


1. A method of treating HIV comprising administering to a patient in need thereof an antibody derivable from a normal subject or from an autoimmune disease subject that binds to a lipid on the surface of HIV or on the surface of HIV-infected cells and thereby neutralizes HIV-1, wherein said antibody is administered in an amount sufficient to effect said treatment.

2. The method according to claim 1 wherein said antibody is derivable from an anti-phospholipid syndrome subject.

3. The method according to claim 1 wherein said antibody is non-pathogenic.

4. The method according to claim 1 wherein said antibody is IS1, IS4 or IS6, or binding fragment thereof.

5. The method according to claim 1 wherein said antibody is IS1, or binding fragment thereof.


again-


Bart Haynes:
"A method of treating HIV comprising administering to a patient in need thereof an antibody
derivable from a normal subject or from an autoimmune disease subject
that binds to a lipid on the surface of HIV
or on the surface of HIV-infected cells
and thereby neutralizes HIV-1"


Bart Haynes:
"A method of treating HIV comprising administering to a patient in need thereof an antibody
derivable from a normal subject or from an autoimmune disease subject
that binds to a lipid on the surface of HIV
or on the surface of HIV-infected cells
and thereby neutralizes HIV-1"


Bart Haynes:
"A method of treating HIV comprising administering to a patient in need thereof an antibody
derivable from a normal subject or from an autoimmune disease subject
that binds to a lipid on the surface of HIV
or on the surface of HIV-infected cells
and thereby neutralizes HIV-1"



------------



It's happening
:)


j

e, obviously because there's more than one paper. That'd be my guess.

and with over 40 people and 9 institutions, I'd say it's likely the case.


Speaking of which, off the top of my head, I'd guess the institutions are, based on already published material & info, in order of my confidence of their involvement;

Duke
UCLA
Harvard
UAB
Fred Hutchinson
SCHARP
Cornell
NIH
Albert Einstein and/or NYU



j

r6,



Regardless of Gendel, realize, Duke has their own PR machine, and they're very good at getting the word out.



j

5. PPHM's / Chen's mab IS1 neutralizes 7 out of 7 HIV strains -------------



This was Haynes work, at the Gates conference, funded with CHAVI money, using Chen's / (PPHM's) mab.


Pojen Chen's IS1 mab has shown some very interesting results. Here is an abstract from the Seattle Gates HIV vaccine conf last fall. Work that was done by Haynes Duke/CHAVI/Gates team, along with - you guessed it- Pojen Chen.


P10-22
Neutralization of HIV-1 primary isolates in PBMC assays
by monoclonal anti-lipid antibodies derived from a
patient with anti-phospholipid syndrome.


MA Moody1, D Montefiori1, MK Plonk1, H Liao1, S Xia1, TC Gurley1,
SM Alam1, P Chen2 and BF Haynes1


1 Duke University Medical Center, Durham, NC, USA;
2 UCLA School of Medicine, Los Angeles, CA, USA


Background: Rare human anti-envelope monoclonal antibodies (Mabs) that
broadly neutralize HIV-1 have been made, but HIV-1+ or vaccinated subjects
rarely make broadly reactive neutralizing antibodies. The observations
that 2F5, 4E10 and 1b12 are polyspecific antibodies and that AIDS may
be rare in patients with primary autoimmune diseases have prompted the
hypothesis that patients with autoimmune diseases who have defects in
tolerance mechanisms may be able to make antibodies that have anti-HIV-1
activity. Thus, we have assayed Mabs derived from anti-phospholipid
syndrome patients for their ability to neutralize HIV-1.

Methods: IS1 and IS4 are human Mabs derived from a patient with antiphospholipid
syndrome. Both are autoantibodies that react with cardiolipin
and phosphatidylserine. IS4 promotes thrombosis in a murine pinch-induced
thrombosis model while IS1 is not prothrombotic and, therefore, is nonpathogenic.
Neutralization assays used were pseudovirus assays in TZM/bl
cells and whole virus assays in peripheral blood mononuclear cells (PBMC).
Interaction of antibodies with Env and lipids was determined by surface
plasmon resonance (SPR) and flow cytometry.

Results: IS1 and IS4 Mabs had no neutralizing activity against HIV-1 primary
isolate pseudoviruses in the TZM/bl assay. In the PBMC assay, however,
both IS1 and IS4 neutralized 7/7 primary isolates tested (Torno, PAVO,
6535, DU123, DU156, DU151, and DU172; IC80s ranged from 0.06-4.1ug/
mL). Against SHIV and SIV viruses IS1 and IS4 neutralized SHIV SP162P3
at IC80s of 0.06 and 0.2μg/mL, respectively, while neither Mab neutralized
SHIV 89,6P or SIVmac239. Using SPR, neither IS1 nor IS4 reacted with
recombinant HIV-1 wild-type Envs. By flow cytometry, neither Mab labeled
viable uninfected T cell lines or PBMC; however, both IS1 and IS4 Mabs
reacted strongly with the surface of virus infected T cells.

Conclusion: Select anti-membrane polyspecific antibodies bind to HIV-1
infected but not uninfected cells, are non-pathogenic, and have anti-HIV-1
neutralizing activity in vitro in PBMC but not pseudovirus assays. It will be
of interest to determine if such non-pathogenic antibodies are protective in
passive therapy trials in vivo in non-human primates. Study of the mechanism
of neutralization of these antibodies and their B cells of origin may provide
clues to novel ways to safely induce protective antibodies to HIV-1.

Supported by the Center for HIV/AIDS Vaccine Immunology AI067854,
CHAVI 005 protocol.

http://www.hivvaccineenterprise.org/_dwn/Late_Breaker_Abstracts.pdf



j

4. UCLA's Pojen Chen's (PPHM's) mabs appear in CHAVI work, under Bart Haynes.....



Pojen Chen and his anti-phospholipid mabs was first mentioned in CHAVI related work in this Sept 2007 CHAVI update:



CHAVI Y03 First Quarter Progress Report-Executive Summary
September 14, 2007
Barton Haynes, MD
CHAVI Director

10. Obtain human monoclonal antibodies from acutely infected HIV-1 patients and from autoimmune disease patients and determine their ability to neutralize HIV-1.

Report: James Robinson, David Montefiori, George Shaw and Lynn Morris comprise our team that is evaluating the specificities of the first neutralizing antibodies produced in acute HIV-1 infection, and have raised monoclonal antibodies that represent these antibodies. Tony Moody and Bart Haynes lead the studies in autoimmune disease patients, and with Pojen Chen at UCLA, have found human mabs from SLE and primary antiphospholipid syndrome patients that broadly neutralize HIV-1 primary isolates.

http://www.chavi.org/modules/chavi_reports/index.php?id=17




j

3. Pojen Chen's team learns insights into antiphospholipid mab binding


In 2006 Chen's group learned WHY they stick to WHAT...

Specifically, why they stick to Beta-2-glycoprotein-I. (because of Arginine in certain places on the mab).

They also learned how to alter the mabs to change their binding specifics, to increase or decrease the mabs attraction to various phospholipids...

For instance, they changed IS4's binding characteristics...


http://www.jimmunol.org/cgi/content/full/177/3/1729



j

jake, I hear ya,

it's also amazing to me since Avastin does almost nothing for overall survival.

but- We haven't knocked the snot out of Avastin- yet....
What we've done is keep pace, and yes- better it, in early results.

If you look at the graphs I posted, you'll see that Avastin drops off a CLIFF! at six months.

If Bavi is still going strong in patients at six months, (and it really is looking great so far), then THAT'S when you can say we're beating Avastin, and the world will take notice.

ALL BAVI PATIENTS AT 4 MONTHS ARE RESPONDERS OR STABLE.

Six months is not far away.

Sure, it's not all the patients in the trial, but if 100% of say, 8 or 10 or 14 patients is progression-free at six months, people will start to see that we've got something very special, unique...


IMHO, the way Bavi works, it should KEEP working..... until?........









-----------

it's happening,

j

2. UCLA's Pojen Chen's anti-PS mabs



Pojen Chen's mabs were originally isolated from an autoimmune disease patient in 1999.

http://www.blackwell-synergy.com/links/doi/10.1111%2Fj.1365-2141.1999.01292.x

Since then, Pojen Chen and others have been doing a lot of experiments with the mabs, ( IS1, IS2, IS4, IS6, CL1 - CL15, etc.), to discern what makes them pathogenic, or non-pathogenic, what are their binding details, etc.




j

1. anybody see this in the 10K yesterday?...................


During June 2007, we entered into an exclusive license agreement with The Regents of the University of California regarding the use of certain Anti-PS antibodies to be used as a possible future generation clinical candidate.


http://www.sec.gov/Archives/edgar/data/704562/000101968708003034/peregrine_10k-043008.htm


So THAT'S why Pojen Chen showed up on Peregrine's SRB back then....

listed under "CORE TECHNOLOGIES"......



CORE TECHNOLOGIES

Pojen P. Chen, Ph.D.
Professor of Medicine, Department of Medicine, Rheumatology Division,
University of California Los Angeles
http://www.peregrineinc.com/index.php?option=com_content&task=view&id=15&Itemid=29


Now it makes sense seeing his name on team Peregrine.

His (Peregrine's) "IS1" mab is pretty interesting.....


j

Duke paper(s) questions from CC -



Questioner:
You also mentioned that the reason for the delay in the Duke paper was that they’re gathering additional data which would make hopefully the results even more exciting. How long will it take to together that additional data?


Steven King:
The work is ongoing. Essentially the work needed for the initial publication and presentations has pretty much been completed. There’s still a little bit of polish up work going, but again these delays are overall a real positive for the program because what we are finding I think is even more exciting than what we originally had hoped going into the end of the studies, and in fact now our effort here internally has probably tripled over the course of the collaboration and the number of outside investigators working with us on the trials, on the studies is also greatly increased. So there is a lot of excitement around what we are doing here, what we are finding, and the potential applications of that knowledge base that we are gathering right now.


Questioner:
So the additional data has been gathered than?


Steven King:
Yes, some of it has. It’s obviously an ongoing process, so the more we learn, the more we get to do research, but as far as the initial publications and what have you, the data is essentially available now. There are publications that are in process and others that have been submitted and so things are moving along very nicely.



---------


j

SK: ANTIVIRAL PROGRAM

continuned collab. w/Duke & others for use in the possible prevention and treatment in HIV

delay has been caused by an expansion of the collab....

currently more than 40 scientists and nine institutions involved.

PL: "There has been a very recent shift in philosophy in NASDAQ" -

'exceptions were rare.
NASDAQ philosophy has now been modified-
the new phil. is to provide co's with the maximum discretion.
as long as a valid definitive plan in place,
- an additional 180 day period with continued listing.
plan must include a plan with a RS,
therefor, if required at that time, we will ask for this.
- at the next SHM.
no plans for a RS yet.
good news,
additional time.'

Interesting. shorts unchanged, still at historic low short interest.



j

DLB NEUTRALIZES INFLUENZA ----------




Peregrine's antiviral DLB --------

Duramycin is a peptide - it's tiny compared to an antibody.

(It can be inhaled, etc...)



The recent PR - (that we're familiar with which I posted here http://investorshub.advfn.com/boards/read_msg.aspx?message_id=30599778 ), mentions the broad US patent grants, and that Thorpe/Soares presented data (at AAI) that the duramycin/biotin conjugate neutralized several enveloped viruses. What they didn't mention specifically, which the below reference does mention, is that the stuff neutralized INFLUENZA.

Duramycin is a peptide - it's tiny compared to an antibody.

It can be inhaled, etc...

-------------------------------------------------

http://www.pharmainfo.ac.cn/view.do?id=5272

duramycin-biotin conjugate (viral infection), University of Texas Southwestern Medical Center/Peregrine Pharmaceuticals

Originator: University of Texas Southwestern Medical Center.

Highest Dev Status: Discovery.

Updated on 14 April 2008.

The University of Texas Southwestern Medical Center (UTSW Medical Center) and Peregrine Pharmaceuticals are investigating duramycin conjugated to biotin, DLB, for the potential treatment of viral infection [ 892975 ]. PRECLINICAL DATA In April 2008, preclinical data were presented at the Experimental Biology meeting in San Diego, CA. In a mouse model of cytomegalovirus (CMV) infection, treatment with DLB (0.8 mg/kg) administered 18 h after infection with CMV followed by daily treatment with DLB was found to increase survival of mice in comparison with those treated with PBS. In vitro, DLB was found to neutralize human influenza A virus in MDCK cells [ 892975 ]. ADDITIONAL INFORMATION By April 2008, the UTSW Medical Center had a research agreement with Peregrine for the development of DLB [ 892975 ]..



---------------------------------------------------
DLB for pulmonary viral infections -------


From Thorpe patent -


[1375] As the earlier study showed duramycin-biotin conjugates to localize to macrophages in the lung following administration in vivo, the stimulation of macrophage-mediated phagocytosis of apoptotic cells shown in the present study has important implications for the therapeutic uses of the present invention, such as in treating pulmonary viral infections.


-----------


j

DLB IN VIVO: 100% Survival -------------



EXAMPLE XXII

PE-Binding Peptide Derivative Treats CMV Infection In Vivo

[1022] In addition to the in vitro anti-viral effects against CMV shown in Example XVII, this example demonstrates that the duramycin-biotin derivative, DLB increased survival of mice infected with mCMV.

[1023] Balb/C mice (6 week old, five mice per group) were infected i.p. with 5.times.10.sup.5 pfu of mCMV RVG102. The mice were treated i.p. on day 1 and every four days with 20 .mu.g/mouse of the duramycin derivative, DLB. Untreated mice served as the control. The mice were monitored for survival past 90 days post infection.

[1024] Treatment with the duramycin-biotin derivative, DLB enhanced survival of the mCMV-infected mice. Mice treated with DLB had 100% survival, as compared to untreated mice, wherein only 25% of the mice survived the infection.


http://appft1.uspto.gov/netacgi/nph-Parser?Sect1=PTO2&Sect2=HITOFF&p=1&u=%2Fnetahtml%2FPTO%2Fsearch-adv.html&r=1&f=G&l=50&d=PG01&S1=%28%22thorpe+philip%22.IN.%29&OS=in/%22thorpe+philip%22&RS=IN/%22thorpe+philip%22



--------


j

DLB virus neutralization in vitro details -------

More from Thorpe patent ---


EXAMPLE XVII

Anti-Viral Effects of PE-Binding Peptide Derivatives

[0933] In addition to the anti-viral effects mediated by anti-PS antibodies, as shown in Example XII and Example XIII, the present example demonstrates the anti-viral effects of peptide derivatives that specifically bind to the other common aminophospholipid, PE.

A. Methods

1. Treatment of CMV-Infected Cells In Vitro

[0934] Confluent monolayers of human diploid foreskin fibroblasts (HHF-R2) in 6-well plates were infected with human CMV AD169 expressing green fluorescent protein (GFP) at an MOI=0.01 as described in Example XII (Bresnahan et al., 1996). The cells were incubated with virus in a total volume of 1.5 ml per well at 37.degree. C. for 90 minutes. During the infection, the plates were gently rocked every 30 minutes. Following the infection, the cell supernatant was removed and DMEM/10% FBS/pen-strep (2 ml per well) was added to each well.

[0935] Different dilutions of duramycin derivatives (DLB).sub.4NA, (DIM).sub.nHIgG, DS-1, DS-2, DS-3 and DC-1 were added to the wells before the addition of the virus, and following infection. The infected cells were incubated at 37.degree. C. for a total of 14 days. The medium and duramycin derivative in each well were replaced every 3 days.

2. Fluorescent Microscopy

[0936] As in Example XII, the recombinant CMV expresses GFP under the control of the SV40 promoter. Hence, infected cells appear green under a fluorescent microscope. In these studies, the CMV-infected cells treated with the duramycin derivatives were observed under a fluorescent microscope at days 4 and 6.

B. Results

[0937] On day 4, there are single infected GFP-positive green cells in untreated wells and wells treated with (DLB).sub.4NA and (DIM).sub.nHIgG. Thus, treatment of HHF-R2 cells with these duramycin derivatives does not appear to inhibit the entry of the virus into the cells. There is some preliminary evidence that the duramycin derivatives DS-1, DS-2 and DS-3 inhibit viral entry into the cells.

[0938] On day 6 after treatment with (DLB).sub.4NA and (DIM).sub.nHIgG, there is a marked difference in the number of infected GFP-positive cells in untreated vs. the duramycin derivative treated wells. By day 6, the virus has spread from the single infected cell seen on day 4 surrounding cells in the untreated wells. However, on day 6 in the wells treated with (DLB).sub.4NA and (DIM).sub.nHIgG, the virus is limited to the original singly infected cell.

[0939] Accordingly, (DLB).sub.4NA and (DIM).sub.nHIgG limit the spread of CMV from the original infected cell to the surrounding cells. This inhibition of viral spread is observed when cells were treated with different concentrations of (DLB).sub.4NA (100 .mu.g/ml and 50 .mu.g/ml) and (DIM).sub.nHIgG (200 .mu.g/ml and 100 .mu.g/ml).

http://appft1.uspto.gov/netacgi/nph-Parser?Sect1=PTO2&Sect2=HITOFF&p=1&u=%2Fnetahtml%2FPTO%2Fsearch-adv.html&r=1&f=G&l=50&d=PG01&S1=%28%22thorpe+philip%22.IN.%29&OS=in/%22thorpe+philip%22&RS=IN/%22thorpe+philip%22


-------


j

DLB for pulmonary viral infections -------


From Thorpe patent -


[1375] As the earlier study showed duramycin-biotin conjugates to localize to macrophages in the lung following administration in vivo, the stimulation of macrophage-mediated phagocytosis of apoptotic cells shown in the present study has important implications for the therapeutic uses of the present invention, such as in treating pulmonary viral infections.


-----------


j

Peregrine's antiviral DLB --------


DLB = duramycin joined to biotin


Duramycin is a peptide - it's tiny compared to an antibody.

(It can be inhaled, etc...)



The recent PR -

(The broad patent grants, and general info about what was presented recently at AAI....)

http://files.shareholder.com/downloads/PPHM/0x0x206630/2a311150-4a0f-494a-8f5f-ffbc7fca5f12/PPHM_News_2008_6_16_Financial.pdf

moby,


Spin this anyway you want, but-

Unless the Bavituximab patients ALL start jumping off a cliff, together, - SOON -

things are looking VERY good for Bavi cancer.










NEXT UP - BAVITUXIMAB VS: LUNG CANCER

(it's already started).


- in patients who have never had chemo.

j

moby,


-


IMO an important difference in the two trials is that in the Avastin trial
21 of the 27 patients had had NO PRIOR CHEMO FOR MBC



As for numbers-

there were 27 patients in the Avastin trial.

0 had a complete response
14 were classified with a partial response
9 were classified with stable disease
2 had progressive disease
2 dropped out

Patients had already started progressing at 2 months


There have (so far) been 14 patients in the Bavituximab trial.

7 have been classified with a partial response
7 have been classified with stable disease.

none of the treated patients have progressed at 2 months,
and none of the patients out 4 months have progressed.








j

moby,


A chart plotting specific results over time is the antithesis of "spin".


j

Progression-Free Survival comparison --------








Bavituximab info:
http://ir.peregrineinc.com/releasedetail.cfm?ReleaseID=319343


Avastin info:
http://clincancerres.aacrjournals.org/cgi/reprint/12/10/3124

(and that comparison is from the earlier phase II "rosey" Avastin trial results, as discussed last Sunday in the NYT in my previous post. Avastin's current breast cancer trial - AVADO, is not seeing similar results.)



------


j