GDVMARCH's AASLD Abstracts 2001- Explanation re: Group 1, Phase II, Hep-B. (l0/l0/01)

You requested my non-scientific layperson's interpretation - DO NOT RELY ON IT -- of Enzo's AASLD 2001 Abstract entitled, "Treatment of Chronic HBV Infection Via Oral Immunemodulation:  Augmentation of the Anti-HBV T-Cell Response and Suppression of Viral Load" (hereinafter called "ABSTRACT #1").  It is scheduled for a Poster presentation in Dallas on November 11, 2001.  It relates to early findings in the not yet completed Group 1 study constituting the first stage of Enzo's Hep-B Phase II human trial in Israel.  It was written to promote discussion among member scientists and not for shareholders or others making market and investment decisions.  The statistics discussed under "RESULTS" are tentative.  They are subject to change upon Group 1 study completion, perhaps around end of this month.  Hence, no intelligent purpose is served by any attempt to evaluate or compare these tentative results with those obtained upon completion of Enzo's Hep-B Phase I.  I used an Online Medical Dictionary at  http://www.graylab.ac.uk/omd/index.html .  Those with relevant scientific backgrounds should feel free to correct.  A copy of ABSTRACT #1 can be accessed for printing and comparison at http://aasld01.agora.com/planner/displayabstract.asp?presentationid=3937.    [My interpretation of the two remaining abstracts will follow in dribs and drabs. This chore is tedious and more suited for one having relevant scientific qualifications.]
 
I INTERPRET ABSTRACT #1, as follows:

BACKGROUND:  Hepatitis B virus ("HBV") -- an unclassified DNA virus with complex, double-layered virions 42nm in diameter and an outer sheath enclosing an inner 27nm core particle containing the circular viral DNA - has three major antigens-namely,  the hepatitis B core antigen (HBcAg), the hepatitis B surface antigen (HbsAg), and hepatitis B e antigen (HbeAg).  HBV is a non-cytopathic virus - meaning, it is a virus that in and of itself does not cause the major disease symptoms in the liver and other cells.  Abstract #1 tells us to look elsewhere other than to the virus for the causes of the resulting hepatocellular (i.e., of or pertaining to liver cells) injury - here, the inflammatory disease leading to swelling and scarring of liver tissues, etc.

Abstract #1 explains the resulting hepatocellular injury is caused or brought about (i.e., mediated) by the inappropriate (i.e., defective or bad) response by the patient's (i.e., the host) own immune system to detection of the existence of the foreign virus and its attempt to fight it.  We have often been told that a person's immune system has a good side (one that helps) and a bad side (one that reacts in a manner that causes more damage than does the disease it is fighting. The latter is what happens when HBV becomes "chronic".  Chronic means the HBV has continued infecting the patient for more than 6-months and/or degenerated into fibrosis, cirrhosis, liver failure, liver cancer and death.

Abstract #1 advises that this excessive bad anti-viral immunity response can be modulated (i.e., lessened or down-regulated).  In this study modulation of the excessive immune response was tried by mouth-feeding the HBV patients certain viral proteins known as antigens - i.e., a concoction of antigens HbsAg+preS1+preS2.

[COMMENT: FYI, antigens are substances which are capable, under appropriate conditions, of inducing a specific immune response and of reacting with the products of that response - i.e., with specific antibodies or specifically sensitized T-lymphocytes, or both.  Antigens describe   anything that is introduced into the body as foreign - e.g., toxins, foreign proteins or particulates, such as bacteria and tissue cells.   An antigen can stimulate the immune system into producing cells that attack it via its polysaccharide molecule (a/k/a antigenic determinant or epitopes) that combines with an antibody or a specific receptor on a lymphocyte (see T-cells below).  Hepatitis B antigens are antigens of the virion of the HBV dane particle, its surface (HbsAg, HBs, Hbe), its core (HBc, HbcAg), and other associated e antigens (HB-Ag, HbeAg).]

[COMMENT: FYI, an antibody is a protein secreted by immune system cells in response to infection. (i.e., A GOOD GUY.) An antibody binds specifically to the BAD GUY antigen and thus stimulates its inactivation by other parts of the immune system. Antibodies to the hepatitis b antigens, include HbsAb to the hepatitis surface antigen
HbsAg, antibody HbcAb to the core of the dane particle (HbcAg), and antibody HbeAb to the "e" antigens Hbe and HbeAg).]

AIMS OF STUDY:  One purpose of this study was to find out what happened to HBV-DNA (i.e., viral load), liver injury (i.e., inflammation, scarring, etc.) and the anti-viral T-cell response in chronic HBV-infected patients while and after they were mouth-fed with HBV envelope* proteins (i.e., * is a lipid membrane packaging a virus particle).

[COMMENT:  (1) Viral load is the amount of virus present in the HBV-infected person's blood stream.  It is usually measured by a polymerase chain reaction ("PCR") quantative test and the result given in number of virus particles per milliliter of blood.  (2)  T-cells are a class of lymphocytes that are derived from and been processed through the thymus. They are primarily involved in controlling cell-mediated immune reactions and in the control of B-cell development.  They also coordinate the immune system by secreting lymphokine hormones.   There are 3 fundamentally different type of T-cells - to wit, helper, killer and suppressor; each has many subdivisions.  T-cells are also called T-lymphocytes.  They bear T-cell antigen receptors (CD3) and lack Fc or C3b receptors.  Major T-cell subsets are CD4 (mainly helper cells) and CD8 (mostly cytotoxic (killer) or suppressor T-cells).  Uncontrolled proliferation of this type of cell gives rise to T-cell leukaemia/lymphoma (e.g., types of cancer).]

METHODS/TESTS EMPLOYED: To find the answers the study sought, its investigators used the following METHODS: They recruited 42 chronic HBV patients who were mouth-fed HBV proteins (HbsAg+preS1+preS2), 3 times a week for 20 to 30 weeks and then followed for another 20 weeks.  During both periods tests were performed on these patients so as to determine what, if any, effect ingestion of the oral formula had on their respective viral load, liver injury, and the response and activity of T-cells.

[COMMENT:  In prior written and teleconference communications Enzo had talked in terms of medicating Group 1, Phase II patients for 30 weeks versus the 20-week period of oral tolerization of Phase I patients.  I called Enzo for clarification of Abstract #1's reference to treatment for 20 to 30 weeks and learned some Phase I patients were included in Group I, Phase II.  They may have required more time or dosing than allowed in Phase I to optimize treatment benefits.  I do not know if former Phase I or new Phase II patients or some of both were treated in Phase II for less than 30 weeks.  Everyone's immune system reacts on a different time schedule.  This was shown by the different number of weeks the assortment of Phase I patients took to register ALT (i.e., alanine aminotransferase), viral load and other related spike-ups and retreats.  Thus, one dosage level or period may not suit all patients.  That's what Phase II and III trials determine.]

Abstract #1 says, patients were monitored throughout for HBV-DNA [viral load] levels, liver enzymes and liver histology.  [COMMENT:  An enzyme is a chemical substance that while remaining unchanged, helps a chemical reaction to take place.  At the other end of the reaction, the enzyme can help in other reactions of the same kind.  The term "liver histology" (a/k/a "microscopic anatomy") refers to biological studies of the minute structure of tissues, cells and organs  -- here, the liver's tissue and cells.]

Also assessed in the laboratory (a/k/a "in vitro") were: (1) the patients' respective HBV-directed T-cell modulations (e.g., investigators observed and determined how the T-cells [i.e., the helper, killer, suppressors] that come into play upon HBV infection were reacting and increasing in numbers and type; plus the effect, if any, they were having on the patient's condition); (2) the cytotoxicity (i.e., an agent or process which is toxic (poisonous) to cells and results in the suppression of cell function or death).  [COMMENT:   E.g., a cytotoxic T-lymphocyte ("CTL") is a lymphocyte that is able to kill foreign cells that have been marked for destruction by the cellular immune system.  It relates to an agent or process that is toxic (poisonous) to cells and results in the suppression of cell function or death.  Cytotoxic T-cells are activated on binding to specific epitopes presented by infected cells or other antigen-presenting cells.  When activated, these CTLs multiply in numbers and attack and kill the diseased cells like an army attacks its enemy.]

The investigators did cytokines assays by using an ELISPOT (i.e., an Elisa immunological assay technique that here identified and measured non-antibody proteins secreted by inflammatory leukocytes and some non-leukocytic cells that act as intercellular mediators).  FWIW, cytokines include the natural interferons, interleukins, tumor necrosis factor, etc.  A RT-PCR (i.e., a Reverse Transcription Polymerase Chain Reaction) technique was used to pick up viral load measurements.

The Phase II patients included in the Abstract #1 results were also followed to determine the number of their peripheral NK T-cells.  [COMMENTS:  NK cells are natural killer T-cells that are sometimes used as a marker.  Their presence in hepatitis-infected individuals appears to be taking on greater importance and will be discussed later in ABSTRACT #2 dealing with colitis and other forms of autoimmune hepatitis.]

ABSTRACT #1 RESULTS: I read the RESULTS of Abstract #1 to mean as follows:  Tests performed during treatment and observation showed: (1) The oral tolerization or mouth feedings of the concoction of abovenamed viral proteins given to the Phase II chronic HBV patients whose results are included in Abstract #1 [NB: we don't know how many of the 42 patients are included therein] elicited a SIGNIFICANT POSITIVE RESPONSE -- i.e., the good part of their immune systems ameliorated a lot of the damage done by the bad part of the patients' immune systems. [THAT'S VERY GOOD and showed again Enzo's scientific principle works.]
 
(2) Specifically, tests done on each patient's blood showed that 46% of participants included in Abstract #1 results had his or her HBV-DNA ("viral loads") decreased [it doesn't say by how much] and that the blood tests also showed 46% of the included patients registered improvement in liver function. [THAT'S A VERY GOOD NUMBER when compared to current treatment efficacy rates.]
 
(3) More accurate liver biopsy tests [i.e., which conduct a microscopic examination of liver tissue removed either during surgery or by insertion of a needle and extraction of sample liver tissue] were performed in the same number of patients.  These tests showed an improvement score of 58% in both hepatitis B surface antigen (HbsAg), and in hepatitis B core antigen (HbcAg). [THAT'S VERY GOOD].
 
(4) In addition, a liver biopsy examination of the minute structure of participants' liver tissues [i.e., the histological test] showed IMPROVEMENT in 33% (~ one-third) of the patients' necroinflammatory [i.e., necrosis is decay or death of liver cells from irreversible inflammatory damage].  [THAT'S VERY GOOD.]
 
(5) Three out of eleven patients who had HbeAg (i.e., Hepatitis B e Antigen) lost the HbeAg.  That meant they were otherwise healthy carriers of antibodies, rather than patients chronically infected by degenerative Hep-B virus. [GOOD because no maintenance needed.]
 
(6) Testing also showed a FAVORABLE (i.e., controlled) INCREASE in specific T-cells that fight HBV. Included was a: (i) 92% increase in T-cells responding to the presence of hepatitis B surface antigen (HbsAg); (ii) 95% increase in cytotoxic T-cell lymphocytes that attack and kill diseased cells; (iii) 71% increase in IFNy-positive (i.e., Interferon, a naturally produced medicine) T-cell clones; and (iv) confirmation that during treatment 80% of the patients whose results are reported in Abstract #1 showed an increase in the number of peripheral NK T-cells (i.e., the type of immune cells which naturally kill infected cells).
 
ABSTRACT #1 CONCLUSIONS are self-explanatory but subject to change because not reflective of treatment results for all Group I, Phase II patients. CONCLUSIONS: Enzo's oral immune-regulation towards HBV-envelope proteins SIGNIFICANTLY AUGMENTED (i.e., produced a substantial increase) in patients' effective anti-viral immunity. It also MARKEDLY DECREASED (i.e., a noteworthy reduction in) viral load, and IMPROVED (enabled healing of) the immune-mediated liver damage (i.e., the injury caused by the bad reaction to the HBV on the part of the untreated immune system).

GDV SUM-UP:  If trial results continue both at or near above efficacy levels and without significant adverse effects, upon regulatory approval Enzo's HBV treatment will be the most effective and least expensive HEP-B treatment of choice worldwide. Enzo =  ~$200/treatment versus others' treatments of low efficacy and @~ $5,000/shot a few times a week for approximately 1 year.  I hope above information helps.