Ronald C. Kennedy

Characterization of anti-hepatitis B surface antigen monoclonal antibodies.

17 monoclonal antibodies generated against purified hepatitis B surface antigen (HBsAg), subtype ayw, were characterized by solid-phase radioimmunoassays. Eleven of these antibodies had specificity against the group-specific alpha determinant of HBsAg, two demonstrated antibody activity against the w HBsAg subtype, one against human serum albumin, and three against human IgG. All monoclonal antibodies were of the IgG class.

A common human anti-hepatitis B surface antigen idiotype is associated with the group a conformation-dependent antigenic determinant.

Abstract Human antibodies that express a common anti-hepatitis B surface antigen idiotype were directed against the group-specific a determinant. The ability of hepatitis B surface antigen (HBsAg)-derived polypeptides to inhibit the idiotype-anti-idiotype reaction was dependent on conformation, since denatured HBsAg viral polypeptides virtually lost their inhibitor capacity when compared to native polypeptides. This report further characterizes a common idiotype associated with human antibodies to HBsAg.

Sequential genital infections by herpes simplex viruses types 1 and 2: Restriction nuclease analyses of viruses from recurrent infections

Virus isolated from a woman presenting with the first symptomatic episode of genital herpes was identified as herpes simplex virus type 1 (HSV-1) by restriction nuclease fingerprinting. Testing for IgM antibody to HSV indicated that the patient had recently contracted a new HSV infection. Virus microneutralization and the micro-solid phase radioimmunometric test for IgG, however, showed that the patient had had prior infection with herpes simplex virus type 2 (HSV-2); thus the HSV-1 infection was acquired despite the presence of antibody to HSV-2. Genital herpes recurred about four, seven, and nine months after the HSV-1 infection. Isolates from the latter three episodes all were of an identical strain of HSV-2 and were not recombinants or a mixture of the viruses. The data show that two distinctly different herpes simplex viruses can initiate genital infections in one individual and suggest that HSV-2 is more likely to recur than HSV-1.

Enhancement of viral hepatitis B antibody (anti-HBs) response to a synthetic cyclic peptide by priming with anti-idiotype antibodies

In vivo injections of anti-idiotype antibodies were used to prime the immune system of mice to hepatitis B surface antigen (HBsAg). Anti-idiotype reagents in conjunction with a cyclic synthetic peptide analogous to positions 122-137 of HBsAg induced an antibody response to HBsAg (anti-HBs) comparable to that obtained with a single injection of intact HBsAg particles. In addition, high anti-HBs titers were produced in mice injected with HBsAg following anti-idiotype priming. These data indicate that anti-idiotype antibodies may be useful in priming the immune system of a host to a potential infectious agent.

Production and characterization of anti-idiotype reagents for the analysis of viral antigen systems

Anti-idiotype reagents have been used recently in the characterization of a number of viral systems. These reagents provide a relatively new approach in viral immunology for the analysis of specific antibody molecules to viruses and their associated antigenic determinants. In this paper we report on the methodology for the generation and characterization of xenogeneic anti-idiotype antibodies in rabbits.

In vivo detection of human hepatoma secreting hepatitis B surface antigen in nude mice with radiolabeled monoclonal antibodies that recognize distinct epitopes

Hepatitis B surface antigen (HBsAg), produced by a human hepatoma which had been transplanted into athymic nude mice, was specifically detected in vivo by 131I-labeled monoclonal antibodies (McAb) directed against distinct epitopes of HBsAg (anti-HBs). Significantly higher levels of radioactivity were present in the hepatoma secreting HBsAg when compared to either a non-HBsAg producing epidermoid tumor or most other tissues obtained from nude mice treated with the 131I-labeled anti-Hbs McAb. A radiolabeled control McAb that did not recognize HBsAg failed to discriminate between either the HBsAg positive and negative tumors or other tissues from nude mice. These data demonstrate the in vivo immunological specificity of anti-HBs McAb for HBsAg associated with a hepatoma tumor.

Hepatitis B Surface Antigen Polypeptide and Synthetic Peptide Vaccines

The hepatitis B vaccine recently licensed for use in the United States has undergone several field trials in high-risk populations and has proven to be immunogenic, efficient and safe. The hepatitis vaccine is unique in that the hepatitis B virus (HBV), the ultimate source of the vaccine, cannot be grown in tissue culture. The immunizing antigen, purified from plasma of healthy human carriers of HBV, consists of noninfectious 22 nm particles of hepatitis B surface antigen (HBsAg), produced as excess viral surface protein and released in the bloodstream. Although the HBsAg particles are highly purified and inactivated, there is some concern that the vaccine may contain traces of host material or infective adventitious agents, which may trigger undesirable effects in the recipients. Ideally, a hepatitis B vaccine candidate should have the following properties: 1) be free of normal human serum protein contaminants, 2) be free of potential residual infectious HBV, 3) be free of other viruses and nucleic acid, 4) have a high degree of immunogenicity when administered in conjunction with an adjuvant suitable for use in humans, 5) be independent of human HBV carriers as a source of immunogenic material, 6) have a reproducible composition, 7) cost less than the present vaccine, and 8) be available in unlimited supplies.

Characteristics of Monoclonal Antibodies to Herpes Simplex Type 2

Monoclonal antibodies were produced by fusing NS-1 cells and spleen cells from BALB/c mice which have been inoculated with either purified herpes simplex virus (HSV) type 2 (HSV-2) virion or the major envelope glycoprotein of HSV-2 with molecular weight 119,000 daltons, designated VP 119. The specificity of each hybridoma antibody has been defined by: 1) western blot techniques employing viral infected cell lysates; 2) virus neutralization (NA); 3) radioimmunoassay (RIA) or; 4) immuno-florescence (FA) using both HSV type 1 (HSV-1) and HSV-2 antigens to determine viral type specificity. Two monoclonal antibodies designated M.186.1B.3 and M.186.2A.2 were produced against HSV-2 virions. One of these (M.186.1B.3) recognized only HSV-2 type specific determinants by RIA and was capable of reducing plaque forming units (PFU) by 70%. The second antibody (M.186.2A.2) recognized both HSV-1 and HSV-2 antigens as determined by either FA or RIA and reduced PFU by 20%. Two monoclonal antibodies designated HS2A and HS5 were generated against VP 119 and detected type specific glycoproteins by RIA. HS2A (anti-VP 119) was further characterized by NA and FA demonstrating partial viral neutralization (20%) and a nuclear staining reaction with HSV-2 infected cells.