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Biomedical subjects

M D Lubeck

Publications and source records attributed to M D Lubeck.

At least 37 records · Page 2Linked to original sources
PubMed1989

Immunogenicity and efficacy testing in chimpanzees of an oral hepatitis B vaccine based on live recombinant adenovirus.

As a major cause of acute and chronic liver disease as well as hepatocellular carcinoma, hepatitis B virus (HBV) continues to pose significant health problems world-wide. Recombinant hepatitis B vaccines based on adenovirus vectors have been developed to address global needs for effective control of hepatitis B infection. Although considerable progress has been made in the construction of recombinant adenoviruses that express large amounts of HBV gene products, preclinical immunogenicity and efficacy testing of candidate vaccines has remained difficult due to the lack of a suitable animal model. We demonstrate here that chimpanzees are susceptible to enteric infection by human adenoviruses type 7 (Ad7) and type 4 (Ad4) following oral administration of live virus. Moreover, after sequential oral immunization with Ad7- and Ad4-vectored vaccines containing the hepatitis B surface antigen (HBsAg) gene, significant antibody responses to HBsAg (anti-HBs) were induced in two chimpanzees. After challenge with heterologous HBV, one chimpanzee was protected from acute hepatitis and the other chimpanzee experienced modified HBV-induced disease. These data demonstrate the feasibility of using orally administered recombinant adenoviruses as a general approach to vaccination.

Adenoviruses, Human↗
PubMed1989

Current use and future directions of adenovirus vaccine.

Adenoviruses are a major cause of respiratory illnesses in military recruits and also are common causes of respiratory and gastrointestinal infections during childhood. Forty-one serotypes of human respiratory and enteric adenoviruses have been identified. Live, oral adenovirus vaccines developed for the military and tested in large clinical trials have proved to be safe and highly effective in decreasing hospitalizations related to adenoviral acute respiratory diseases. Studies have demonstrated little horizontal transmission among military personnel but substantial transmission among family members. Use of recombinant techniques have opened new opportunities for the development of recombinant adenovirus vector vaccines against a number of viral pathogens such as hepatitis B, human immunodeficiency, herpes simplex and respiratory syncytial virus.

Adenoviridae↗
PubMed1988

A new hamster model for adenoviral vaccination.

Both adult and baby hamsters infected intranasally with human adenovirus type 5 exhibited virologic, serologic, and histologic evidence of infection. When 8-day old hamsters were infected with 4 x 10(6) pfu, concentrations of virus up to 2 x 10(6) pfu/animal were detected in the lung, peaking on day 2. The minimum infectious dose was 1 x 10(3) pfu/animal. This model may be useful in studies of conventional and recombinant adenoviral vaccines for humans.

Adenoviridae Infections↗
PubMed1988

Killing of human tumor cell lines by human monocytes and murine monoclonal antibodies.

We evaluated the capacity of freshly isolated blood monocytes to mediate antibody-dependent cellular-mediated cytotoxicity (ADCC) in cooperation with murine anti-tumor monoclonal antibodies (MAbs). Blood monocytes isolated from most donors by adherence selection to fibronectin-coated plastic surfaces and subsequently depleted of natural killer/killer (NK/K) cells exhibited significant ADCC activity against tumor cell lines in combination with an IgG3 antitumor MAb (BR55-2). However, significant variation in ADCC competence was observed among donors. Culture parameters influencing monocyte ADCC activity were evaluated and optimized. The influence of MAb isotype on ADCC capacity of anti-tumor MAbs was also evaluated using anti-tumor class-switch variant hybridoma proteins and a panel of anti-tumor MAbs. MAbs of the IgG2a and IgG3 subclasses exhibited high ADCC potential, whereas MAbs of the IgG2b subclass exhibited no ADCC activity. One of two IgG1 MAbs tested exhibited high ADCC activity with monocyte effectors. The role of monocytes or macrophages in tumor remission of cancer patients undergoing MAb immunotherapy is not known. However, correlative studies of monocyte ADCC capacity and responsiveness of cancer patients to MAb immunotherapy may help to establish the role of these effectors in MAb-mediated tumor remissions.

Animals↗
PubMed1988

Expression of HBV surface antigen or HIV envelope protein using recombinant adenovirus vectors.

Recombinant adenoviruses were constructed that contained either the HBsAg coding sequence or the HIV envelope protein coding sequence. The recombinant adenoviruses can replicate normally in cultured human cells. Cells infected with the adenovirus-HBV recombinant secreted HBsAg into the tissue culture medium. This HBsAg had immunological and physical properties similar to those of the 22-nm particles found in human serum. Expression of HIV envelope protein in cells infected with the adenovirus-HIV recombinant was demonstrated using cytoimmunofluorescence and immunoprecipitation. A hamster model was developed to evaluate the immunogenic properties of adenovirus-HBV recombinants. Hamsters inoculated intranasally with live adenovirus-HBV recombinant produced antibody against both adenovirus and hepatitis B virus surface antigen.

Adenoviridae↗
PubMed1987

Monoclonal antibody-defined correlations in melanoma between levels of GD2 and GD3 antigens and antibody-mediated cytotoxicity.

A monoclonal antibody is described that specifically detects the ganglioside antigens GD2 and GD3, binding preferentially to GD2, in melanoma. Antibody specificity was demonstrated with solid-phase radioimmunoassay and enzyme-linked immunosorbent assay as well as by immunostaining on thin-layer chromatography plates using structurally characterized gangliosides. Binding of both the IgG3 antibody and its IgG2a switch variant were assayed on live cells by cytofluorography and by immunoperoxidase staining on frozen tissue sections. The binding patterns correlated with antitumor activity in antibody-dependent cellular cytotoxicity and complement-dependent cytotoxicity assays with human effector cells and complement in an 111In-release assay using cell lines derived from the same individual. The significant level of killing in all tumor cells tested that express GD2, GD3, or both, suggests the importance of multiple specificity towards tumor antigens, i.e., binding of a monoclonal antibody to two or more tumor-associated antigens.

Animals↗
PubMed1987

Recombinant adenovirus induces antibody response to hepatitis B virus surface antigen in hamsters.

Recombinant adenoviruses carrying the hepatitis B virus surface antigen coding sequence in the adenovirus E3 region were constructed using DNA from either adenovirus type 5 or an adenovirus type 5 E3-region deletion mutant. Both of these recombinant adenoviruses replicated as efficiently as wild-type adenovirus in all human cells tested, including the human diploid cell strain WI-38. This indicates that insertion of the hepatitis B virus surface antigen gene into the E3 region does not significantly affect viral replication. Human cells infected with these recombinant adenoviruses secreted immunoreactive hepatitis B virus surface antigen. Since a practical small animal model for human adenoviruses was lacking, a hamster model was developed to evaluate the immunogenic potential of these recombinant adenoviruses. Upon intranasal inoculation, both wild-type adenovirus and the adenovirus E3-region deletion mutant replicated in the lungs of these animals and induced an antibody response against adenovirus. Hamsters similarly immunized with the live recombinant adenoviruses produced antibody against both adenovirus and hepatitis B virus surface antigen.

Adenoviruses, Human↗
PubMed1985

Isolation and characterization of anti-monosialoganglioside monoclonal antibody 19-9 class-switch variants.

Three different monoclonal antibody isotypes, IgG1, IgG2b, and IgG2a, were derived by selection of isotype-switch variants from the CO-19-9 hybridoma. All three antibodies retained their binding specificities and affinities and bound to the same epitope--as defined by the anti-idiotype analysis. Availability of gamma 1, gamma 2b, and gamma 2a Ig heavy chain variants directed against the same epitope on the monosialoganglioside antigen permitted detailed analysis of their Fc fragment receptor (FcR) binding affinities, their cytolytic activities (antibody-dependent, macrophage-mediated cytotoxicity) in vitro, and their tumoricidal activities in vivo. Analysis of the binding of three isotypes to the human FcR expressed by U937 cells induced by gamma interferon has shown that only IgG2a proteins bound to high-affinity FcR, but not IgG1 or IgG2b variants. Although all three isotypes were active in the antibody-dependent, macrophage-mediated cytotoxicity assay with murine thioglycolate-elicited macrophages, the IgG2a gave the highest percentage of lysis; similar results were obtained in the same assay with human monocytes as effector cells. In the nude mice experiment, only the IgG2a variant inhibited growth of human colorectal carcinoma, while IgG1 and IgG2b were ineffective. Thus, selection of isotype-switch variants resulted in the conversion of monoclonal antibody from noncytolytic to cytolytic with possible immunotherapeutic application.

Animals↗
PubMed1985

The interaction of murine IgG subclass proteins with human monocyte Fc receptors.

The use of murine monoclonal antibodies in the immunotherapy of human disease has prompted interest in the interactions of murine IgG with Fc receptors (FcR) expressed on human effector cells. We examined the heterocytophilic interactions between monomeric murine IgG subclass proteins and the FcR expressed on human monocytic cells (peripheral blood monocytes and interferon (IFN)-gamma-induced U937 cells). All four murine IgG2a antibodies and both murine IgG3 antibodies that were tested bound to human monocyte FcR with high affinity (10(8) to 10(9) M-1). By contrast, the affinities of four murine IgG1 and four IgG2b monomers were 100-fold to 1000-fold lower than the affinity of the human IgG1-FcR interaction. A 68,000 to 72,000 dalton protein was isolated by affinity chromatography from blood monocytes and from IFN-gamma-induced U937 cells on murine IgG2a, IgG3, and human IgG immunoadsorbents. In binding assays with IFN-stimulated U937 cells, murine IgG2a and IgG3 antibodies showed complete cross-blocking with a human IgG1 myeloma protein, indicating that murine and human IgG interact with the same population of Fc-binding proteins. No evidence for heterogeneity of cross-reactive FcR was observed. The ability of murine IgG2a and IgG3 monomers to compete with human IgG1 monomers for binding to human monocyte FcR suggests the potential usefulness of antibodies of these isotypes in the immunotherapy of diseases in which monocyte- or macrophage-mediated, antibody-dependent cellular cytotoxicity may play a role in the modification or remission of disease.

Animals↗
PubMed1984

Induction of mouse IgG2a- and IgG3-dependent cellular cytotoxicity in human monocytic cells (U937) by immune interferon.

The effects of natural and recombinant human gamma-interferon (IFN-gamma) on mouse monoclonal antibody-dependent cellular cytotoxicity (ADCC) mediated by U937 human monocytic-like cells were examined. The efficiency of mouse monoclonal antibody of different isotypes in inducing ADCC was also compared. The number of receptors for the Fc portion of immunoglobulin G (IgG) (FcR) for mouse IgG2a and IgG3 on U937 cells, as detected by IgG antibody-sensitized erythrocyte rosette formation, was significantly enhanced by IFN-gamma. In contrast, FcR for mouse IgG1 and IgG2b were not detected even after IFN-gamma stimulation. U937 cell-mediated ADCC against sheep or ox red blood cell targets was minimal. However, after incubation with human purified IFN-gamma, U937 cells exhibited increased activity in IgG2a- and IgG3-dependent lysis, whereas their activity in IgG1- and IgG2b-dependent lysis was low. ADCC stimulated by IFN-gamma was inhibited by Protein A. When mouse peritoneal exudate cells were used, FcR for all IgG isotypes were easily detected, and all IgG isotypes mediated ADCC. Taken together, these results indicate that IFN-gamma induces U937 cell ADCC with mouse IgG2a and IgG3 partly through augmentation of FcR expression. Recombinant IFN-gamma showed the same effect as natural IFN-gamma. These effects of IFN-gamma were completely abrogated by anti-IFN-gamma serum but not by anti-IFN-alpha or normal rabbit serum. Addition of polymyxin B or lipopolysaccharide did not affect the activity of IFN-gamma.

Animals↗
PubMed1983

Human macrophages armed with murine immunoglobulin G2a antibodies to tumors destroy human cancer cells.

Macrophages isolated from tumor-bearing patients as well as cultured human monocytes express Fc receptors that cross-react strongly with murine immunoglobulins of the G2a but only slightly or not at all with the G1, G2b, or G3 subclasses. Such macrophages in the presence of murine immunoglobulin G2a monoclonal antibodies to tumors mediated the killing of tumor cells in vitro. These data suggest that monoclonal antibodies of the G2a subclass may be useful in the immunotherapy of human cancer.

Animals↗
PubMed1979

Adenovirus-associated virus structural protein sequence homology.

Adenovirus-associated virus (AAV) structural proteins (VP1, VP2, and VP3) have been examined to determine if areas of sequence homology exist between these three virion proteins. Tryptic and chymotryptic maps have been produced which demonstrate extensive areas of sequence homology common to all three proteins. The amino acid compositions of the proteins were also determined and were found to be very similar. These data are consistent with the hypothesis that all three virion proteins arise either from a common precursor of similar transcripts.

Amino Acid Sequence↗
PubMed1978

Susceptibility of influenza A viruses to amantadine is influenced by the gene coding for M protein.

Influenza A virus recombinants derived from "resistant" and "sensitive" parental viruses were examined for susceptibility to inhibition by amantadine. Correlation of gene constellation and amantadine susceptibility revealed that the gene coding for M protein influences sensitivity or resistance to amantadine. All recombinants which derived an M protein from an amantadine-resistant parent were found to be resistant to amantadine. All amantadine-sensitive recombinants derived an M gene from the amantadine-sensitive parent. However, a few amantadine-resistant recombinants which derived an M gene from the sensitive parent were also isolated, suggesting that the expression of amantadine sensitivity in these recombinants may be influenced by other genes.

Amantadine↗