The HERV-K102 – AFP Immunosenescence Paradigm for Functional Cures for HIV-1MARIAN LADEROUTE, PH.D. MEDICAL SCIENCES – IMMUNOLOGYIMMUNE SYSTEM MANAGEMENT CLINIC & LAB (VOLUNTARY CONSULTANT)

OTTAWA, ONTARIO CANADAEmail: hervk102@bell.net Skype:

hervk102

The new paradigm and evidence supporting it is covered in detail in:

Laderoute MP. A new paradigm about HERV-K102 particle production and blocked release to explain cortisol mediated immunosenescence and age-associated risk of chronic disease. Discovery Medicine 20(112):379-391, 2015.

Human Endogenous Retrovirus –K102

(HERV-K102) HERV-K102 is a fully functional and replication competent endogenous retrovirus found in HIV-1 patients (first characterization of such, Laderoute M et al., AIDS 2007, Open AIDS Journal, 2015)

HERV-K102 is also the first foamy retrovirus of humans discovered (Laderoute M et al., AIDS 2007, Open AIDS Journal, 2015)

HERV-K HML-2 group members including HERV-K102, are the most active and most intact HERVs, where expression has been associated with many diseases

Accumulating evidence shows HERV-K102 or similar antigen expression is protective against HIV-1 replication and in breast cancers

Human Endogenous Retrovirus –K102

(HERV-K102) Particle production in HIV-1 infection has been shown by two groups: Laderoute M et al., AIDS 2007 and Contreras-Galindo R et al., J.Virol. 2012

Particles can reach 10 11 per ml of plasma in 84 hours, so a very rapid induction is possible

Particle production has been implicated in resistance to HIV-1 acquisition (Laderoute M et al., Open AIDS Journal, 2015)

However, HERV-K102 particle production is about 7 logs inhibited in HIV-1 patients (a mean of about 8,200 particles per ml of plasma) when compared with patients with other bloodborne pathogens

About 96 % of HIV-1 patients have detectable HERV-K102 particles (cDNA) and/or Env antibodies to HERV-K102 specific epitopes. Incidence and levels of antibodies seem higher in HIV-1 patients than patients with other blood borne pathogens (Laderoute MP et al., 2007)

HERV-K102 particle production generates foamy macrophages

BackgroundImmunosenescence (immune aging) refers to a dysregulated immune system associated with aging. It comprises both immunosuppression and concomitant inflammation presumably at the level of macrophages. Macrophages orchestrate innate and adaptive arms of immunity.

Immunosenescence is thought to be caused by increased cortisol and decreased dehydroepiandrosterone (DHEA) levels associated with aging and persistent infections.

How macrophages display both immunosuppression and inflammation concurrently, has remained an enigma.

As well, how this relates to increase risk of cancer, infectious diseases, autoimmunity etc., has not been elucidated.

Finally its role in HIV-1 pathogenesis remained unclear.

Background- AFPSince the mid 70’s, alpha-fetoprotein (AFP) is a well established (but largely ignored) immunosuppressive factor which also confers apoptosis (cell death) resistance by binding to the 67 kD AFP receptor (AFPr).

This AFPr is found on macrophages (Laderoute & Pilarski, 1994) and is overexpressed on common adenocarcinomas (Laderoute MP, Ph.D Thesis 1991, Laderoute et al, 1994).

Increased levels of AFP are prognostic for progression for a number of viral infections ( Zhu M et al., 2015; Cheng J et al., 2014; and reviewed in Mizejewski GJ, 2001; Terentiev AA and Moldogazieva, 2013).

DHEA and flavonoids block the induction and bioactivity of AFP.

Background- AFPAFP may also bind to CCR5 on primary macrophages through its carbohydrates and blocks binding by R5-HIV-1 strains (Atemezem A et al., 2002; Seddiki N et al., 1997) raising the possibility of a role of active AFP in the switch from CCR5 to CXCR4 co-receptor usage with HIV-1 progression.

This binding to CCR5 which is also expressed on Tregs (de Oliveira CE et al., 2014) may help explain how AFP may contribute to non-antigen specific T suppressor T cells (Murgita RA et al., 1977, Gershwin ME et al., 1978) now known as Tregs.

AFP levels directly correlate with HIV-1 viral loads (Gross S et al., 2003).

Proposed Key Roles of the Alteration of the DHEA/Cortisol Ratio in Immunosenescence and HIV-1

1. Laderoute MP. A new paradigm about HERV-K102 particle production and blocked release to explain cortisol mediated immunosenescence and age-associated risk of chronic disease. Discovery Medicine 20 (112): 379-391, 2015.

DHEA is considered an anti-stress/youth hormone and levels diminish with aging. DHEA (but not DHEA-S), appears to bind active AFP and renders it inactive.1

As well, cortisol induces AFP expression.

Accordingly as you age or with HIV-1 infection, your levels of active AFP in the system may increase due to diminished DHEA and increased cortisol.

This may be relevant to immunosenescence (Deeks SG et al., 2012) and the diminished DHEA/cortisol ratio as found with progression in HIV-1 patients (Clerici M et al., 2000).

It is likely HIV-1 induces AFP expression in macrophages but needs to be confirmed.

Ordinarily in normal healthy adults, a viral infection induces HERV-K102 particle production in macrophages rendering them foamy as HERV-K102 is a foamy virus. On day 7, the foamy macrophages lyse, releasing the HERV-K102 particles which then can antagonize HIV-1 replication. In aged or unhealthy adults, there is insufficient DHEA and/or flavonoids in the system to counterbalance cortisol and AFP. Active AFP blocks the lytic release of HERV-K102 particles in HIV-1 infected cells and could be induced by HIV-1. The potent protection against HIV-1 replication by HERV-K102 particles and T and B cell autoimmunity is lost when there is too much active AFP in the system. The loss of DHEA/insufficient flavonoids contributes to progression.

How might HERV-K102 protect the host? Our understanding of HERV-K102 host protection mechanisms has been best studied in HIV-1 pathogenesis. Three of the four proposed mechanisms are expected to apply to tumors and other viruses or intracellular pathogens.

Four Postulated Mechanisms for HERV-K102 Antagonism Against HIV-1

Model for HIVHIV AntagonismAntagonismby HERVHERV--K102K102

Non-InfectiousHIV Released

Adaptive Immune Adaptive Immune System ActivationSystem Activation

HERV-K 102 Induced

LysisLysis

Lysis Lysis ofof HIV Infected CellsHIV Infected Cellsbyby Anti-HIV T cells/

Antibody

HIV

HERV-K102 Particles Released

22

33

LysisLysis of HIV Infected Cells byHERVHERV--K102 ParticlesK102 Particles

Lysis Lysis of HIV of HIV **infected cells byinfected cells by

Anti-HERV-K102 T cells/

Antibody

4

11

From: Laderoute MP, Discovery Medicine, 2015.

1. Molecular Antagonism2. Lysis of HIV-infected

Cell Producing HERV-K102 Particles

3. Lytic Infection of Abnormal Cells (oncolytic and virolytic) and Increased Proviral Copy Number in Normal Cells (arming)

4. Expansion of Autoimmune T and B Cells to HERV-K Antigens (TLR mediated?), the latter which Behave as Surrogate Antigens for Targeting Transformed Cells

Part 1: proposed HERV-K102 –AFP Immunosenescence Paradigm in Elite Controllers

(EC) HIV-1 induces both AFP expression/secretion and HERV-K102 particle production in

macrophages (can also be induced/enhanced by other intracellular infections and by cortisol)

Where sufficient DHEA and/or flavonoids exists in the host (as may be the case in EC), the bioactivity of AFP is inhibited, allowing the lytic release of HERV-K102 particles on day 7 from foamy macrophages

HERV-K102 particles are thought to be virolytic which is enhanced when active AFP in the system is blocked by DHEA and/or flavonoids

These particles may generate “autoreactive” T and B cell responses to HERV-K antigens expressed only on virus-infected cells or tumors, but which are not found at the cell surface of normal cells (temporary “innate vaccine” possibly involving TLR signalling). Antibodies to HERV-K102 envelope (Type 1 HML-2 epitopes) may directly mediate apoptosis.

Part 2: proposed HERV-K102 –AFP Immunosenescence Paradigm in HIV-1

Progressors When DHEA and/or flavonoid levels are not sufficient (with age for

example or following chronic stress, chronic infection, poor nutrition etc., ), AFP binds to the 67 kD AFPr on macrophages and blocks the lytic release of HERV-K102 particles

This causes foamy macrophages to linger, which is well known to initiate atherosclerosis and increase the risk of cardiovascular disease

As well, these dysfunctional macrophages are not able to activate adaptive immunity, while active AFP in the system blocks afferent and efferent immunity

The oncolytic and virolytic activity of HERV-K102 particles is lost as : a) particles are not released, which prevents activation of autoimmune T and B cell responses and b) activity of the particles is blocked by AFP preventing the lysis of virally infected cells

HERV-K102 envelope or similar antigens are not expressed on the surface of normal cells but are found as cell surface beacons of tumor transformed or virally infected cells for T and B cell autoimmune reactivity. This has been shown to mediate clearance of HIV-1 infected cells (and breast cancer cells). For example T cell activity against HERV-K102 envelope was found in EC and cleared HIV-1 infected cells in vitro (SenGupta D et al., 2011; Jones RB et al., 2012).

HERV-K102 particle production in vivo has been associated with mediating remissions in limited studies of CFS, MS and an acute Epstein barr virus infection (Laderoute et al, 2007).

In the case of breast cancer, HERV-K102 envelope was surprisingly shown to directly mediate apoptosis when triggered by antibody, both in vitro and in vivo. (Wang-Johanning F et al, 2012).

So what does the HERV-K102 –AFP immunosenescence paradigm say about HIV-1 functional cures?

HERV-K102 is a naturally occurring oncolytic/virolytic virus, and particles behave as “innate vaccine” so one gets both when particles are administered (or induced and released from foamy macrophages).

CAVEAT: But you must include measures to inactivate AFP in the system for this to work.

How to Exploit HERV-K102 particles to enhance levels of Post-Interruption

Controllers (PIC)Administer orally, optimal levels of DHEA, flavonoids and amino acids (the latter to support HERV-K102 particle production, for example high lysine (K) is needed) allowing a 3-4 week lead time before ART treatment interruption:

Administer agents able to induce HERV-K102 particle production (such as cortisol for 5 days) and/or inject HERV-K102 particles, and/or

Administer monoclonal antibodies to HERV-K102 envelope protein (which have been shown to directly mediate apoptosis of target cells)

Follow progress (before, during lead time and after) by doing qPCR on plasma samples quantitate: 1) HERV-K102 particle production (cDNA), 2) HERV-K102 integration in genomic DNA sloughed into plasma, 3) HIV-1 viral loads, and 4) integrated levels of HIV-1 in genomic DNA sloughed into plasma

Optionally, monitor active AFP levels (see Lester EP et al., 1976)

How to utilize HERV-K102 for HIV-1 prevention

(i.e., innate vaccines for short term protection*).Administer orally, optimal levels of DHEA, flavonoids and amino acids

(the latter to support HERV-K102 particle production, for example high lysine (K) is needed) :

ACTIVE innate immunization: Administer agents able to induce HERV-K102 particle production (such as cortisol for 5 days), and/or inject HERV-K102 particles, and/or

PASSIVE innate immunization: Administer monoclonal antibodies to HERV-K102 type 1 envelope protein epitopes (which have been shown to directly mediate apoptosis of target cells)

Do not co-administer traditional vaccines within 3-6 month windows of each other due to cross-interference

* After high risk behaviour or when an infant is born to an HIV-1 positive mother.

How to utilize HERV-K102 for HIV-1 prevention

and sterilizing controlResearch and Develop a recombinant sterilizing vaccine by using HERV-K102 as the vector which contains nucleotides able to block HIV-1 replication (and not HERV-K102) such as ant-Tat (Scarborough RJ et al., 2015, Cafaro A et al., 2015). Conduct Safety Evaluation in BLT mice. Administer orally the conditioning regime [optimal levels of DHEA, flavonoids and amino acids (the latter to support HERV-K102 particle production, for example high lysine (K) is needed)] allowing a 3-4 week lead time: Administer vaccine orally or by parenteral injection. Administer cortisol for 5 days. Continue conditioning regime. Test uptake by quantitation of HERV-K102 and recombinant HERV-K102 cDNA in

plasma (cDNA) and ratio of recombinant to normal in genomic DNA sloughed into plasma. Determine optimal plasma ratio to be associated with protection against HIV-1 acquisition.

Repeat process until ratios are sufficient to ensure protection.

Note: Do not co-administer traditional vaccines within 3-6 month windows of each other due to cross-interference

Caveats on using HERV-K102 antigens for traditional vaccine approaches

T and B cell autoimmune responses are only temporary and are suspected to not involve traditional T cell help. Therefore breaking tolerance is an issue.

At the present time, using HERV-K102 particles (induction or administration) may be the safest way to induce T and B cell “autoimmunity” to HERV-K HML-2 antigens, but protection is only expected to last about 6-12 months.

There is still the need to ensure adequate DHEA and flavonoids in the host to enable or facilitate HERV-K102 (proposed) virolytic activity and/or antibody ability to clear tumors by blocking apoptosis resistance mediated by AFP.

HERV-K102 is unique to humans and is not easily modelled in animal models. As well, cortisol does not induce AFP expression in rodents indicating even humanized BLT mice may not be perfect for pre-clinical testing.

Conclusions: Immunosenescence appears to involve HERV-K102 particle production and blocked release by AFP in foamy macrophages, the latter which then linger and initiate atherosclerosis and other diseases related to immunosenescence.

HIV-1 is known to involve immunosenescence and a decreased DHEA/cortisol ratio is associated with progression.

HERV-K102 particle production and/or Env specific antibody occurs in about 96 % of HIV-1 patients and AFP levels correlate with HIV-1 viral loads.

HERV-K102 particles (induction or administration) may be useful as a virolytic virus therapy or innate vaccine for HIV-1 provided sufficient DHEA, flavonoids (and amino acids) are ensured in the host. It may be difficult to test HERV-K102 in animal models.

HERV-K102 may be a preferred vector for recombinant sterilizing cures against HIV-1 as it naturally occurs in humans while the antibody response it generates may also naturally induce apoptosis in HIV-1 infected cells, while not reactive with normal cells.

The discovery of human endogenous retrovirus K102 (HERV-K102) as a protector foamy virus of humans has been patented world wide by the Public Health Agency of Canada (National Microbiology Lab, Winnipeg, MB Canada), for which inventors receive no benefits.

Dr. Laderoute retired from Immune System Management Clinic & Lab in 2015 during the writing of the new HERV-K102 –AFP immunosenescence paradigm, but remains as a voluntary consultant (since 1998).

Conflict of Interest Statement