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M J Reddehase

Publications and source records attributed to M J Reddehase.

At least 55 records · Page 3Linked to original sources
PubMed1991

Protection against lethal cytomegalovirus infection by a recombinant vaccine containing a single nonameric T-cell epitope.

The regulatory immediate-early (IE) protein pp89 of murine cytomegalovirus induces CD8+ T lymphocytes that protect against lethal murine cytomegalovirus infection. The IE1 epitope is the only epitope of pp89 that is recognized by BALB/c cytolytic T lymphocytes (CTL). Using synthetic peptides, the optimal and minimal antigenic sequences of the IE1 epitope have been defined. To evaluate the predictive value of data obtained with synthetic peptides, recombinant vaccines encoding this single T-cell epitope were constructed using as a vector the hepatitis B virus core antigen encoded in recombinant vaccinia virus. In infected cells expressing the chimeric proteins, only IE1 epitope sequences that were recognized as synthetic peptides at concentrations lower than 10(-6) M were presented to CTL. Vaccination of mice with the recombinant vaccinia virus that encoded a chimeric protein carrying the optimal 9-amino-acid IE1 epitope sequence elicited CD8+ T lymphocytes with antiviral activity and, furthermore, protected against lethal disease. The results thus show for the first time that recombinant vaccines containing a single foreign nonameric CTL epitope can induce T-lymphocyte-mediated protective immunity.

Amino Acid Sequence↗
PubMed1990

Distinct gamma/delta T cell receptors define two subsets of circulating porcine CD2-CD4-CD8- T lymphocytes.

The species swine provides the only example for CD2+ and CD2- subsets of Ig-CD4-CD8- lymphocytes with the propensity for homing to lymphoid tissue (Saalmüller et al., Eur. J. Immunol. 1989. 19: 2011). That the CD2-CD4-CD8- lymphocytes are bare of marker molecules that typify T lymphocytes raised the question of whether or not this cell type is descended from the T lymphocyte differentiation lineage. It is documented that expression of a phylogenetically conserved external epitope of T cell receptor gamma/delta subdivides porcine CD2- lymphocytes into an epitope 86D+ minor and an 86D- major subset. Expression of distinct forms of the T cell receptor gamma/delta, disulfide-bonded N-glycosylated surface heterodimers of under reducing conditions 38/40 and 37/40 kDa, respectively, hallmarks the CD2-86D+ and CD2-86D- subsets both as T lymphocytes.

Animals↗
PubMed1990

Porcine gamma/delta T lymphocyte subsets differing in their propensity to home to lymphoid tissue.

Three types of T cell receptors (TcR) gamma/delta were identified in swine, all of which are disulfide-bonded cell surface heterodimers. The first, a 38/40-kDa molecule, is related to the TcR gamma/delta of sheep and cattle, and defines in swine a subset of CD4-CD8- T lymphocytes that is enriched in the circulating T lymphocyte pool. The other two receptors are private to swine. The second, a 37/40-kDa molecule, is expressed by the major subset of CD2-CD4-CD8- T lymphocytes that is neither enriched in nor excluded from lymphoid tissue. By contrast, the third type of TcR gamma/delta, a 40/46-kDa molecule, is found on CD2+CD4-CD8- T lymphocytes accumulating in lymphoid tissue. Thus, TcR gamma/delta T lymphocyte subsets differing in homing propensity are typified by the expression of distinct molecular forms of the TcR gamma/delta.

Animals↗
PubMed1989

Structure of the gene of tum- transplantation antigen P91A: the mutated exon encodes a peptide recognized with Ld by cytolytic T cells.

Mutagen treatment of mouse P815 tumor cells produces immunogenic mutants that express new transplantation antigens (tum- antigens) recognized by cytolytic T cells. We found that the gene conferring expression of tum- antigen P91A contains 12 exons, encoding a 60 kd protein lacking a typical N-terminal signal sequence. The sequence shows no significant similarity with sequences in current data bases. A mutation that causes expression of the antigen is located in exon 4; it is the only apparent difference between the normal and the antigenic alleles. A short synthetic peptide corresponding to a region of exon 4 located around this mutation makes P815 cells sensitive to lysis by anti-P91A cytolytic T cells. The mutation creates a strong aggretope enabling the peptide to bind the H-2 Ld molecule. Several secondary tumor cell variants that no longer express tum- antigen P91A were found to carry deletions in the gene.

Amino Acid Sequence↗
PubMed1989

Presentation of CMV immediate-early antigen to cytolytic T lymphocytes is selectively prevented by viral genes expressed in the early phase.

The regulation of antigen processing and presentation to MHC class I-restricted cytolytic T lymphocytes was studied in cells infected with murine cytomegalovirus. Recognition by cytolytic T lymphocytes of the phosphoprotein pp89, the immunodominant viral antigen expressed in the immediate-early phase of infection, was selectively prevented during the subsequent expression of viral early genes. The surface expression of MHC class I glycoproteins and their capacity to present externally added pp89-derived antigenic peptides were not affected. Because recognition of several other antigens occurred during the early phase, a general failure in processing and presentation was excluded. Since neither rate of synthesis, amount, stability, nor nuclear transport of pp89 was modified, the failure in recognition indicates a selective interference with pp89 antigen processing and presentation.

Animals↗
PubMed1989

Site-restricted persistent cytomegalovirus infection after selective long-term depletion of CD4+ T lymphocytes.

We have established a murine model system for exploring the ability of a CD4 subset-deficient host to cope with cytomegalovirus infection, and reported three findings. First, an antiviral response of the CD8 subset of T lymphocytes could be not only initiated but also maintained for a long period of time despite a continued absence of the CD4 subset, whereas the production of antiviral antibody proved strictly dependent upon help provided by the CD4 subset. Second, no function in the defense against infection could be ascribed as yet to CD4-CD8- T lymphocytes, which were seen to accumulate to a new subset as a result of depletion of the CD4 subset. This newly arising subset did not substitute for CD4+ T lymphocytes in providing help to B lymphocytes, and was also not effective in controlling the spread of virus in host tissues. As long as a function of these cells in the generation and maintenance of a CD8 subset-mediated response is not disproved, caution is indicated with concern to an autonomy of the CD8 subset. Third, even though with delay, the CD8+ effector cells raised in the CD4 subset-deficient host were able of clear vital tissues from productive infection and to restrict asymptomatic, persistent infection to acinar glandular epithelial cells in salivary gland tissue.

Animals↗
PubMed1989

A pentapeptide as minimal antigenic determinant for MHC class I-restricted T lymphocytes.

Peptides that are antigenic for T lymphocytes are ligands for two receptors, the class I or II glycoproteins that are encoded by genes in the major histocompatibility complex, and the idiotypic alpha/beta chain T-cell antigen receptor. That a peptide must bind to an MHC molecule to interact with a T-cell antigen receptor is the molecular basis of the MHC restriction of antigen-recognition by T lymphocytes. In such a trimolecular interaction the amino-acid sequence of the peptide must specify the contact with both receptors: agretope residues bind to the MHC receptor and epitope residues bind to the T-cell antigen receptor. From a compilation of known antigenic peptides, two algorithms have been proposed to predict antigenic sites in proteins. One algorithm uses linear motifs in the sequence, whereas the other considers peptide conformation and predicts antigenicity for amphipathic alpha-helices. We report here that a systematic delimitation of an antigenic site precisely identifies a predicted pentapeptide motif as the minimal antigenic determinant presented by a class I MHC molecule and recognized by a cytolytic T lymphocyte clone.

Amino Acid Sequence↗
PubMed1989

Phenotypic discrimination between thymic and extrathymic CD4-CD8- and CD4+CD8+ porcine T lymphocytes.

The composition of the T lymphocyte population in swine is special in that in addition to the CD4-CD8+ subpopulations, CD4-CD8- and CD4+CD8- and CD4+CD8+ subpopulations are prominent in the peripheral circulating as well as in the resident T lymphocyte pools. Since the same phenotypes are characteristic of thymic populations, it was asked whether the unusual distribution in swine may result from an emigration of thymic precursor phenotypes to the periphery. This explanation was refuted, as all thymic subpopulations were found to express CD1, albeit with differences in antigen density, whereas all extrathymic subpopulations lack CD1. The cellular distribution of CD2 in swine is without precedent among all species studied. Whereas in sheep and cattle the extrathymic CD4-CD8- subpopulation is known to entirely lack CD2 and to have a low propensity for homing to lymphoid tissues, the CD4-CD8- subpopulation in swine splits into CD2+ and CD2- subsets, both of which do reside in lymphoid tissues. While CD2+CD4-CD8- T lymphocytes are rare in the circulating pool, this subset accumulates in spleen and lymph nodes. This may indicate a role for CD2 in homing. Thus the species swine is immunologically unique, not only because of having CD1-CD2+CD4+CD8+ T lymphocytes in the periphery, but also with regard to subdivision and homing behavior of its CD4-CD8- T lymphocyte subpopulation.

Animals↗
PubMed1988

Failure in generating hemopoietic stem cells is the primary cause of death from cytomegalovirus disease in the immunocompromised host.

We have shown in a murine model system for cytomegalovirus (CMV) disease in the immunocompromised host that CMV infection interferes with the earliest detectable step in hemopoiesis, the generation of the stem cell CFU-S-I, and thereby prevents the autoreconstitution of bone marrow after sublethal irradiation. The antihemopoietic effect could not be ascribed to a direct infection of stem cells. The failure in hemopoiesis was prevented by adoptive transfer of antiviral CD8+ T lymphocytes and could be overcome by syngeneic bone marrow transplantation. CD8+ T lymphocytes and bone marrow cells both mediated survival, although only CD8+ T lymphocytes were able to limit virus multiplication in host tissues. We concluded that not the cytopathic effect of virus replication in host tissues, but the failure in hemopoiesis, is the primary cause of death in murine CMV disease.

Animals↗
PubMed1988

Molecular basis for cytolytic T-lymphocyte recognition of the murine cytomegalovirus immediate-early protein pp89.

The murine cytomegalovirus protein pp89, which is encoded by gene ieI, is a nonstructural regulatory protein expressed in the immediate-early phase of the viral replication cycle and located mainly in the nucleus of infected cells. Protection of BALB/c (H-2d) mice against a lethal murine cytomegalovirus challenge infection is achieved by vaccination with a recombinant vaccinia virus, MCMV-ieI-VAC, expressing pp89 as the only murine cytomegalovirus gene product. The protection is entirely mediated by T lymphocytes of the CD8+ subset. In the present report, we analyzed the molecular basis of the recognition of pp89 by BALB/c CD8+ cytolytic T lymphocytes. A series of internal and terminal deletion mutants of gene ieI was constructed and cloned in vaccinia virus, and the antigenicity and immunogenicity of the fragments of pp89 expressed by the recombinants were studied. A region of only one-sixth of the protein, from amino acids 154 to 249 and encoded by the fourth exon of gene ieI, was sufficient for both the recognition in vitro of the protein by pp89-specific cytotoxic T lymphocytes and the induction in vivo of pp89-specific cytotoxic T lymphocytes. By using synthetic peptides, the sequence between residues 161 and 179, which is located within the defined domain, was identified as an epitope presented to BALB/C cytotoxic T lymphocytes by the class I major histocompatibility antigen Ld.

Amino Acid Sequence↗
PubMed1988

Adoptive immunotherapy of murine cytomegalovirus adrenalitis in the immunocompromised host: CD4-helper-independent antiviral function of CD8-positive memory T lymphocytes derived from latently infected donors.

The ability of memory T lymphocytes derived from latently infected mice to control murine cytomegalovirus disease in the immunocompromised host was studied by adoptive transfer experiments. At a stage of pathogenesis when virus had already colonized target tissues, a therapeutic antiviral function could be ascribed to the CD8+ subset. This in vivo function was not restricted to sites in which intravenously infused lymphocytes usually are trapped or home in, such as the lungs or the spleen, respectively, but was also evident in the adrenal glands, a site to which antiviral effector cells have to specifically migrate. Specific infiltration of adrenal gland cortical tissue by donor-derived CD8+ memory T lymphocytes was demonstrated. CD4+ memory T lymphocytes had no antiviral effect by themselves and also were not required for the function of the CD8+ effector cells in this short-term immunotherapy model. These findings should help settle the debate about which subset of T lymphocytes comprises the effector cells that can directly control cytomegalovirus infection in the murine model system.

Adrenal Gland Diseases↗
PubMed1988

A nonstructural viral protein expressed by a recombinant vaccinia virus protects against lethal cytomegalovirus infection.

The nonstructural immediate-early protein pp89 of murine cytomegalovirus (MCMV) is the first viral protein synthesized after infection and has a regulatory function in viral gene expression. Despite its localization in the nucleus of infected cells, pp89 is also the dominant antigen recognized by MCMV-specific cytolytic T lymphocytes. The recombinant vaccinia virus MCMV-ieI-VAC, which expresses pp89, was used to study the capacity of this protein to induce protective immunity in BALB/c mice. Vaccination with MCMV-ieI-VAC induced a long-lasting immunity that protected mice against challenge with a lethal dose of MCMV but did not prevent infection and morbidity. In vivo depletion of CD8+ T lymphocytes before challenge completely abrogated the protective immunity. CD8+ T lymphocytes derived from MCMV-ieI-VAC-primed donors and adoptively transferred into sublethally irradiated and MCMV-infected recipients were found to limit viral replication in host tissues, whereas CD4+ T lymphocytes and pp89-specific antiserum had no protective effect. The data demonstrate for the first time that a single nonstructural viral protein can confer protection against a lethal cytolytic infection and that this immunity is entirely mediated by the CD8+ subpopulation of T lymphocytes.

Animals↗
PubMed1988

Immune system of swine: dissection of mononuclear leucocyte subpopulations by means of two-colour cytofluorometric analysis.

Expression of the differentiation antigen 8/1, a porcine leucocyte marker for which no counterpart is known as yet in other species, was tested in relation to the expression of surface Ig and the monocyte marker 74-22-15. B lymphocytes and T lymphocytes can be typified by the exclusive expression of surface Ig and 8/1, respectively, while all monocytes defined by the expression of the 74-22-15 antigen co-express 8/1 with the highest staining density observed for this marker. 8/1+ T lymphocyte subsets defined by the expression of CD4 and CD8 cell surface antigens differed in the expression of 8/1 with the staining density being high in CD4+ T lymphocytes, intermediate in CD8+ as well as in CD4+CD8+ T lymphocytes, and low in the CD4-CD8- subset. Practically all mononuclear leucocytes present in peripheral blood are included in the subpopulations defined by this kit of reagents, and thus it can be predicted that if mononuclear leucocytes which do not belong to any of these subpopulations exist at all, these must be rare and will not be accessible to an analysis without enrichment.

Animals↗
PubMed1987

A nonstructural polypeptide encoded by immediate-early transcription unit 1 of murine cytomegalovirus is recognized by cytolytic T lymphocytes.

We have constructed target cells by cotransfection of the MHC gene Ld and fragments of murine cytomegalovirus (MCMV) DNA coding for nonstructural immediate-early (IE) proteins. Transfectants were tested by using CTL clone IE1 with specificity for an IE epitope presented in association with Ld. Data show that clone IE1 recognizes a product of the ie1 transcription unit of MCMV, and that its specificity is shared by approximately 25% of polyclonal IE-specific CTL. The results provide the first definite evidence that expression of a herpes virus IE gene encoding a regulatory protein gives rise to antigen expression detectable by specific CTL.

Animals↗