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Y Thanavala

Publications and source records attributed to Y Thanavala.

12 recordsLinked to original sources
PubMed2001

Oral immunization with hepatitis B surface antigen expressed in transgenic plants.

Oral immunogenicity of recombinant hepatitis B surface antigen (HBsAg) derived from yeast (purified product) or in transgenic potatoes (uncooked unprocessed sample) was compared. An oral adjuvant, cholera toxin, was used to increase immune responses. Transgenic plant material containing HBsAg was the superior means of both inducing a primary immune response and priming the mice to respond to a subsequent parenteral injection of HBsAg. Electron microscopy of transgenic plant samples revealed evidence that the HBsAg accumulated intracellularly; we conclude that natural bioencapsulation of the antigen may provide protection from degradation in the digestive tract until plant cell degradation occurs near an immune effector site in the gut. The correlate of protection from hepatitis B virus infection is serum antibody titers induced by vaccination; the protective level in humans is 10 milliunits/ml or greater. Mice fed HBsAg-transgenic potatoes produced HBsAg-specific serum antibodies that exceeded the protective level and, on parenteral boosting, generated a strong long-lasting secondary antibody response. We have also shown the effectiveness of oral delivery by using a parenteral prime-oral boost immunization schedule. The demonstrated success of oral immunization for hepatitis B virus with an "edible vaccine" provides a strategy for contributing a means to achieve global immunization for hepatitis B prevention and eradication.

Animals↗
PubMed2000

Production of hepatitis B surface antigen in transgenic plants for oral immunization.

Here we present data showing oral immunogenicity of recombinant hepatitis B surface antigen (HBsAg) in preclinical animal trials. Mice fed transgenic HBsAg potato tubers showed a primary immune response (increases in HBsAg-specific serum antibody) that could be greatly boosted by intraperitoneal delivery of a single subimmunogenic dose of commercial HBsAg vaccine, indicating that plants expressing HBsAg in edible tissues may be a new means for oral hepatitis B immunization. However, attainment of such a goal will require higher HBsAg expression than was observed for the potatoes used in this study. We conducted a systematic analysis of factors influencing the accumulation of HBsAg in transgenic potato, including 5' and 3' flanking elements and protein targeting within plant cells. The most striking improvements resulted from (1) alternative polyadenylation signals, and (2) fusion proteins containing targeting signals designed to enhance integration or retention of HBsAg in the endoplasmic reticulum (ER) of plant cells.

Administration, Oral↗
PubMed1999

Immunization of mice with P6 of nontypeable Haemophilus influenzae: kinetics of the antibody response and IgG subclasses.

The kinetics of the anti-P6 antibody response was characterized in three strains of mice of different haplotypes (Balb/c; H-2d, C3H/H; H-2k, SJL/J; H-2s). Anti-P6 antibodies were measured on a weekly basis by enzyme-linked immunosorbent assay (ELISA). The primary response peaked 2 or 3 weeks after the initial injection with 40 microg of purified P6. The response remained at a plateau for 8-10 weeks. A maximum titer of 1:1,638,400 was attained and then steadily declined. To study the ability of P6 to generate a recall response, we opted to boost the vaccinated mice with a known subimmunogenic dose of live nontypeable Haemophilus influenzae (NTHI) bacteria. After the anti-P6 antibody titers in the primed animals had stayed at baseline levels for 2 weeks, the mice were injected intraperitonealy with 10(8) cfu of NTHI in sterile saline. This challenge with live NTHI bacteria induced a very rapid and strong secondary antibody response in all mice. Finally, we demonstrated that these murine anti-P6 sera were 100% bactericidal against three strains of NTHI when tested in a complement dependant bactericidal assay.

Animals↗
PubMed1998

Evaluation of B and T-cell responses in chimpanzees immunized with Hepagene, a hepatitis B vaccine containing pre-S1, pre-S2 gene products.

Approximately 5-10% of healthy young adults receiving the commercially available hepatitis B vaccine (either serum derived or recombinant) fail to mount an adequate immune response. This nonresponder rate has prompted the demand for more immunogenic vaccines. An alternative to the currently licensed hepatitis B vaccines is Hepagene, a novel recombinant hepatitis B vaccine containing S, pre-S1 and pre-S2 antigenic components, produced in the mouse C127I clonal cell line after transfection of the cells with genes encoding the three antigens. In this study, chimpanzees were immunized with Hepagene to study the humoral and cellular immune responses to this vaccine. Two out of the three animals immunized with this vaccine seroconverted 4 weeks after their first injection and all of the animals elicited high anti-HBs levels that were maintained for at least 28-30 weeks after their third immunization. The anti-HBs levels elicited in these animals protected them against an experimental challenge with HBV. Peripheral blood mononuclear cells (PBMCs) obtained from immunized animals could be stimulated in vitro by rHBsAg and peptides representing regions within all three of the viral envelope proteins. Additionally, an anti-id that mimics the a determinant in the S-region of HBsAg could also stimulate in vitro proliferation of PBMCs from these immune animals. These results indicate that this new recombinant HBV vaccine encoding all three of the surface antigen proteins is highly immunogenic is that it can stimulate strong cellular and humoral immune responses.

Animals↗
PubMed1996

Novel approaches to vaccine development against HBV.

This review summarizes studies from my laboratory based on two novel approaches towards vaccine development against hepatitis B surface antigen. The first approach deals with the ability of a monoclonal anti-idiotypic antibody to functionally mimic the a determinant of HBsAg. The second approach exploits the strategy of genetically engineering plants so that they can be used as inexpensive alternatives to fermentation systems for the production of candidate vaccine antigens.

Amino Acid Sequence↗
PubMed1995

Immunogenicity of transgenic plant-derived hepatitis B surface antigen.

The focus of the Children's Vaccine Initiative is to encourage the discovery of technology that will make vaccines more readily available to developing countries. Our strategy has been to genetically engineer plants so that they can be used as inexpensive alternatives to fermentation systems for production of subunit antigens. In this paper we report on the immunological response elicited in vivo by using recombinant hepatitis B surface antigen (rHBsAg) purified from transgenic tobacco leaves. The anti-hepatitis B response to the tobacco-derived rHBsAg was qualitatively similar to that obtained by immunizing mice with yeast-derived rHBsAg (commercial vaccine). Additionally, T cells obtained from mice primed with the tobacco-derived rHBsAg could be stimulated in vitro by the tobacco-derived rHBsAg, yeast-derived rHBsAg, and by a synthetic peptide that represents part of the a determinant located in the S region (139-147) of HBsAg. Further support for the integrity of the T-cell epitope of the tobacco-derived rHBsAg was obtained by testing the ability of the primed T cells to proliferate in vitro after stimulation with a monoclonal anti-idiotype and an anti-idiotype-derived peptide, both of which mimic the group-specific a determinant of HBsAg. In total, we have conclusively demonstrated that both B- and T-cell epitopes of HBsAg are preserved when the antigen is expressed in a transgenic plant.

Amino Acid Sequence↗
PubMed1995

Delineation of the minimal hepatitis B surface antigen-specific B- and T-cell epitope contained within an anti-idiotype-derived pentadecapeptide.

A pentadecapeptide (2F10 peptide) is capable of mimicking the group-specific "a" determinant of human hepatitis B surface antigen (HBsAg) at both the B- and the T-cell level. This peptide represents a sequence on the heavy-chain hypervariable region of a monoclonal "internal image" anti-idiotype (anti-id 2F10) that has partial sequence homology to the "a" determinant epitope of HBsAg. To identify the exact location of the B- and T-cell epitopes, four truncated peptides (peptides 1-4) were synthesized. Using these truncated peptides we have identified the minimal sequence (octapeptide 3) that represents a functional B- and T-cell epitope capable of generating HBsAg-specific antibodies and T cells. This to our knowledge represents the first example of a short peptide sequence functioning as both a B- and a T-cell epitope. We have also identified another T-cell epitope (2F10 peptide 4), but this peptide fails to elicit HBsAg-specific B cells and T cells. Thus, the 2F10 pentadecapeptide is composed of two nonoverlapping, functional T-cell epitopes only one of which is HBsAg specific. Since peptide 3 represents the complementarity-determining region and peptide 4 represents the framework region of the anti-id 2F10, we conclude that an 8-aa sequence from the complementarity-determining region of anti-id 2F10 is sufficient for the molecular mimicry of HBsAg. Finally, our experiments suggest that sequences flanking the minimal immunodominant epitope exert a considerable influence on the nature of antigenic processing that occurs and the resultant T-cell reactivity elicited.

Amino Acid Sequence↗
PubMed1995

A comparison of the antibody and T cell response elicited by internal image and noninternal image anti-idiotypes.

We have previously reported that several monoclonal anti-idiotypes were generated against a syngeneic monoclonal antibody designated H3F5 (idiotype) which recognizes the protective "a"-determinant on hepatitis B surface antigen (HBsAg). In the present study, we compared the humoral and cellular response to HBsAg by immunizing Balb/C mice with four of these anti-Ids. We observed that two internal image and one noninternal image anti-Ids can act as surrogates for HBsAg. Following immunization with these anti-Ids, high levels of anti-HBs antibodies were elicited. Three of the anti-Ids could also prime T cell for an HBsAg-specific proliferative response, the two internal image anti-Ids elicited much stronger T cell responses to HBsAg than the non-internal image anti-Id. One noninternal image anti-Id did not generate any anti-HBs antibody response and could not elicit an HBsAg-specific T cell proliferative response. All four anti-Ids could elicit immunoglobulin which was H3F5-idiotype positive. This study further supports the concept that some monoclonal anti-Ids can mimic at both the B and T cell level the properties of a nominal antigenic epitope.

Animals↗
PubMed1993

Mimicry of the a determinant of hepatitis B surface antigen by an antiidiotypic antibody. I. Evaluation in hepatitis B surface antigen responder and nonresponder strains.

B and T cell responses of several strains of mice, immunized with a monoclonal antiidiotype (anti-Id) that mimics the a determinant of hepatitis B surface antigen (HBsAg), were studied to determine if the immune response to the anti-Id was regulated by H-2-linked immune response genes as has been previously observed for HBsAg. We report that immunization with anti-Id could elicit HBsAg-specific antibodies in mice of the H-2d,q, or f haplotype and in an outbred wild mouse strain (Mus spretus), thus circumventing the H-2 haplotype restriction pattern observed when immunizing with HBsAg in H-2f mice. Purified lymph node T cells from mice of the H-2d or q haplotype and M. spretus that were primed in vivo with HBsAg or anti-Id could be stimulated in vitro with either HBsAg or anti-Id but not with an irrelevant antibody of the same subclass as the anti-Id. However, purified lymph node T cells from H-2f mice that were primed in vivo with the anti-Id could only be stimulated in vitro with anti-Id. No in vitro stimulation whatsoever was observed in H-2f mice immunized with HBsAg. The effect of processing and presentation of the anti-Id by antigen-presenting cells (APC) was studied in mice of the H-2d haplotype. Stimulation of purified lymph node T cells by HBsAg and anti-Id was shown to be strictly dependent on APC and restricted by major histocompatibility complex class II antigens at the I-A locus. Treatment of APC with paraformaldehyde or chloroquine abrogated the T cell response to all antigens except for a nine-amino acid synthetic peptide representing a partial analogue of the group a determinant of HBsAg S(139-147). The significance of these results is discussed in the context of understanding the mechanism of mimicry elicited by the anti-Id.

Amino Acid Sequence↗
PubMed1992

Molecular mimicry of hepatitis B surface antigen by an anti-idiotype-derived synthetic peptide.

Monoclonal antibody 2F10 is an "internal-image" anti-idiotype (anti-id) antibody capable of mimicking the group-specific "a" determinant of human hepatitis B surface antigen (HBsAg). By mRNA sequencing and computer-assisted molecular modeling of monoclonal antibody 2F10, we identified a 15-amino acid region of the heavy-chain hypervariable region that has partial residue homology with sequences of the "a" determinant epitopes of HBsAg. We have established that a linear 15-mer peptide from a contiguous region on the anti-id antibody can (i) generate anti-HBsAg-specific antibodies when injected into mice, (ii) prime murine lymph node cells for in vitro HBsAg-specific T-cell proliferative responses, and (iii) stimulate in vitro human CD4+ T cells that were primed in vivo to HBsAg by natural infection with hepatitis B virus or vaccination with a commercially available HBsAg vaccine. Significantly, this peptide could also stimulate CD4+ T cells of human hepatitis B virus carriers. We conclude that a 15-mer peptide derived from the anti-id sequence can duplicate the B- and T-cell stimulatory activity of the intact anti-id antibody and the antigen that is mimicked, HBsAg.

Amino Acid Sequence↗