Search PubMed⌕ Search

Biomedical subjects

J Reimann

Publications and source records attributed to J Reimann.

At least 19 recordsLinked to original sources
PubMed1992

Immunization with soluble simian virus 40 large T antigen induces a specific response of CD3+ CD4- CD8+ cytotoxic T lymphocytes in mice.

C57BL/6 (B6) mice (H-2b) were immunized with the large tumor antigen (T Ag) of simian virus 40 (SV40). Intraperitoneal or subcutaneous sensitization with soluble T Ag specifically primed cytotoxic lymphocyte precursors (CTLp). T Ag-specific cytotoxic T lymphocytes (CTL) were detected in a cytotoxicity assay after specific in vitro restimulation of effector cell populations from mice immunized with 2-10 micrograms purified, soluble T Ag and boosted with an injection of 2 micrograms T Ag 2-4 weeks after priming. Cells used for in vitro restimulation and as targets in cytotoxicity assays were syngeneic (B6-derived) RBL5 lymphoma cells expressing SV40 T Ag after transfection with a T Ag-encoding expression vector. Effector cells of this response were H-2 class I-restricted CD3+ CD4-CD8+ CTL. The magnitude of the anti-T Ag CTL response of B6 mice stimulated by soluble virus protein was comparable to the anti-T Ag CTL response of SV40-infected B6 mice. Injections of denatured or native T Ag protein primed CTLp equally well, but immunization with an equal dose of antigen emulsified in incomplete Freund's adjuvants inefficiently stimulated CTLp.

Animals↗
PubMed1992

Bone marrow cells of athymic nude mice express functional T cell receptor alpha chain transcripts rearranged to V delta 2, 3, 4, 5, 6 genes.

From bone marrow cells (BMC) of athymic nude mice, T cell receptor (TcR) alpha chain transcripts were selectively amplified by polymerase chain reaction (PCR) using V delta 2-, V delta 3-, V delta 4-, V delta 5-, V delta 6- and C alpha-specific primers. Amplified DNA fragments were cloned, and 32 randomly selected clones from 5 PCR were sequenced. Twenty-three distinct rearrangement events were detected, of which 87% (20/23) were in-frame. All five tested V delta genes (V delta 2, 3, 4, 5, 6) rearranged in-frame to J alpha-C alpha. N-region diversity in V delta-J alpha junctions present in most clones was limited to two to five nucleotides. P-nucleotide additions in this region were also detected. The V delta 5 gene located 3' of C delta in reversed transcriptional orientation was rearranged to J alpha by inversion. The J alpha usage pattern of the sequenced clones was strongly biased towards rearrangement of the most 5' genes (located nearest to C delta) of the J alpha cluster: the most 5' J alpha (J alpha TA1) was used by 30% of all clones, and 78% of all J alpha rearranged to V delta were located in the 5' 12 kb of the 60-kb J alpha cluster. As distinct V delta/C delta and V alpha/C alpha TcR usage patterns are prevalent in peripheral T cell populations, our data suggest that these TcR usage patterns results from repertoire selections operating in alpha beta and gamma delta T cell lineages, but not from preferential V delta-C delta and V alpha-C alpha rearrangement patterns.

Animals↗
PubMed1992

H-2 class I Ld antigen positively selects different TCR V beta gene products in CD8+ and CD4+ T cells.

Expression of CD4+ or CD8+ determinants and ten different TCR V beta genes was analysed by two-colour flow cytometry in lymph node cells (LNC) of BALB/c and dm2 (a BALB/c Ld loss mutant) mice. TCR V beta 14 expression of CD8+ T cells and TCR V beta 7 and 14 of CD4+ T Cells were significantly more frequent in BALB/c than in dm2 LNC, whereas no differences were observed in the frequency distribution of TCR V beta+ CD4-CD8- double negative LNC from BALB/c versus dm2 mice. These results represent the first published evidence for positive selection of particular TCR V beta genes by individual MHC class I molecules in non-manipulated mice.

Animals↗
PubMed1992

[Reposition and suture fixation of dislocated posterior chamber lenses].

We treated 11 patients with posterior chamber intraocular lenses that were dislocated into the vitreous cavity. Using a pars plana approach, vitrectomy was performed and in order to facilitate manipulation, half of the bulbus volume was filled by perfluorodecaline. The IOL was grasped with forceps and repositioned with a special technique. In this way the first haptic is fixed in the ciliary sulcus. Fixation of the second haptic with 9 x 0 polypropylene suture was performed with a similar procedure. Finally, prophylactic cryotherapy of the retina is indicated.

Eye Foreign Bodies↗
PubMed1992

CD3+ T cells in severe combined immunodeficiency (scid) mice. VI. Rescue of scid-derived, IgM-producing B cells by transfer of CD4+ CD8- T cells from various lymphoid organs.

Intravenous injection of purified CD4+ CD8- T cells from thymus, spleen or lymph nodes of adult dm2 donor mice (H-2d, Ld-, IgMa) into young C.B-17 scid/scid (scid) mice (H-2d, Ld+, IgMb) partially and selectively reconstituted the splenic CD4+ T-cell compartment of recipient scid mice. This was demonstrated by cytofluorographic analyses, histological examinations, growing donor-derived CD4+ T-cell lines in vitro from spleens of transplanted scid mice, and serial passage of Ld-, CD3+CD4+CD8- T-cell lines through young scid recipients for more than 1 year. Host-derived IgMb appeared in sera of all CD4+ T-cell-transplanted young scid mice, while none of the sex- and age-matched, non-transplanted control scid mice was Ig+. B-cell leakiness was most efficiently induced by transfer of low numbers of adult CD4+ CD8- thymocytes: transfer of 10(3) purified CD4+ CD8- thymocytes efficiently rescued scid-derived B cells, while transfer of 2 x 10(7) non-fractionated thymocytes from the same donor mouse was inefficient. Serum antibodies of scid mice with T-cell-induced B-cell leakiness stained congenic 'double-positive' (CD4+ CD8+) thymocytes and immunoprecipitated protein bands with an apparent MW of 14,000, 28,000, 43,000, 65,000 and 80,000 from 125I-labelled thymocytes. These data indicate that repopulation of peripheral lymphoid organs with CD4+ T cells is always accompanied by partial co-reconstitution of the B-cell system, and raises the question of the interdependence of these two lymphocyte subsets.

Animals↗
PubMed1991

CD3+ T cells in severe combined immune deficiency (scid) mice. I. Transferred purified CD4+ T cells, but not CD8+ T cells are engrafted in the spleen of congenic scid mice.

Young (less than 3 months of age) and old (greater than 1 year of age) C.B-17 scid/scid mice were tested for the presence of immunoglobulin in serum and CD3+ T cells in spleen and peritoneal cavity. In all old severe combined immune deficiency (scid) mice tested we found detectable, but very variable levels of serum immunoglobulin as well as splenic and peritoneal CD3+ T cells comprising 3% to 10% of the nonfractionated cell populations of these organs (n = 10). In contrast, none of the analyzed young scid mice showed any evidence of peripheral lymphocytes. Low numbers (2 x 10(5) to 5 x 10(5) cells/mouse) of highly purified CD4+ cells from congenic C.B-17 or BALB/c donor mice were injected intravenously into young scid recipient mice. A CD4+ T cell population was clearly engrafted when transplanted scid mice were analyzed 8 to 13 weeks after T cell transfer: (a) a CD3+CD4+CD8- T cell population was detectable in the spleens of all recipient scid mice by flow microfluorometry analyses; (b) CD3+CD4+CD8 T cell lines could be grown out of these spleens in vitro; (c) the histological examination revealed evidence of lymphoid cell repopulation in the spleens of all transplanted scid mice and (d) transplanted CD4+ T cell populations could be serially transferred into secondary and tertiary recipient scid mice. These data indicate that scid mice can be constructed in which only the CD4+ T cell compartment is selectively reconstituted. In contrast to the successful engraftment of CD4+ T cell, highly purified congenic CD8+ T cells could not be engrafted into the spleen of scid mice.

Aging↗
PubMed1991

CD3+ T cells in severe combined immunodeficiency (scid) mice. II. Transplantation of dm2 lymphoid cells into semi-allogeneic scid mice.

Intravenous injection of nonfractionated BALB/c-H-2dm2 (dm2) (Ld-) spleen cells into 4-week-old, semi-allogeneic (H-2d, Ld+) C.B-17 scid/scid severe combined immunodeficient (scid) mice (2 x 10(7) cells/mouse) reconstituted T lymphopoiesis in thymi and repopulated the lymphoid white pulp in spleens of these immunodeficient recipients. Transplantation of dm2 thymocytes into young scid mice (5 x 10(7) cells/mouse) established a donor-derived CD3+ T cell population in spleens of recipient scid mice, in which CD4+T cells predominated. This was demonstrated by marker analyses of thymocytes and splenocytes, and determinations of serum immunoglobulin levels in transplanted scid mice. Transfer of splenocytes from young primary scid recipients into young secondary or tertiary recipients (3 x 10(6) cells/mouse) engrafted preferentially dm2-derived CD3+CD4+CD8- T cells in spleens of scid mice despite the strong selective Ld-associated alloantigenic stimulus for CD8+ T cells. Intravenous injections of nonfractionated dm2 spleen cells (2 x 10(7) cells/mouse) or thymocytes 5 x 10(7) cells/mouse) into 10- to 12-month-old, "leaky" scid mice induced severe clinical signs of graft-vs.-host disease (GVHD) in all scid recipients. Lymphoid repopulation of spleen and thymus in old scid recipients was incomplete. This GVHD was not transferrable by injecting 3 x 10(6) spleen cells from old diseased primary scid recipients into secondary or tertiary young scid recipient mice. In these serial transfers, dm2-derived CD3+CD4+CD8- T cells were again preferentially engrafted in spleens of scid recipients. Transfer of purified CD4+ dm2 T cells into young scid mice (2 x 10(5) to 5 x 10(5) cells/mouse) engrafted this T cell subset into the spleen of semi-allogeneic scid recipients. This was revealed by histological examinations, surface marker analyses, in vitro isolation of donor-type CD3+CD4+ T cell lines from spleens of transplanted scid mice, and serial transfer experiments. These data indicated that the CD4+ T cell compartment of scid mice can be selectively repopulated by semi-allogeneic T cells. Injections of purified CD8+ dm2 T-cells into young scid mice (2 x 10(5) cells/mouse) did not establish a CD8+ T cell graft in spleens of recipients. It was necessary to inject transplanted scid mice biweekly with 10(4) units recombinant interleukin 2 to establish and/or maintain transferred dm2 CD8+ T cells in spleens of these recipients, dm2 CD8+ T cell-transplanted and interleukin 2-treated scid mice did not develop any evidence of GVHD over the 9-week observation period.

Age Factors↗
PubMed1991

CD3+ T cells in severe combined immunodeficiency (scid) mice. IV. Graft-vs.-host resistance of H-2d scid mice to intravenous injection of allogeneic H-2b (C57BL/6) spleen cells.

Intraperitoneal injection of 2 x 10(7) nonfractionated spleen cells (SC) from C57BL/6 (B6, H-2b) mice into completely allogeneic immunodeficient H-2d scid mice induced clinical and histological signs of acute graft-vs.-host disease (GVHD), with all transplanted severe combined immunodeficiency (scid) mice dying in the 3rd week post-transfer. In contrast four out of five scid mice survived for greater than 7 weeks after intravenous (i.v.) injections of equal numbers of B6 SC. Intravenously allotransplanted scid mice analyzed in the 8th week post-transfer had engrafted donor-type CD4+ and CD8+ T cells in the spleens but showed no clinical or histological evidence of GVHD. i.v. injection of 10(7) or 10(6)O B6 SC engrafted allogeneic T cells in spleens of scid recipients; in contrast, i.v. injection of 10(5) nonfractionated B6 SC or 3 x 10(5) cell sorter-purified, naive or anti-H-2d-primed splenic CD4+ or CD8+ B6 T cells led to rejection by young scid recipient mice. B6 T cells engrafted into spleens of scid mice after i.v. injection showed proliferative anti-host alloreactivity in vitro. No cytotoxic reactivity against host-type alloantigens was found in standard 4-h 51Cr-release assays. These data demonstrate that allogeneic T cells injected i.v. into immunodeficient scid mice are partially tolerized against host-type alloantigens.

Animals↗
PubMed1991

CD3+ T-cells in severe combined immunodeficiency (scid) mice. III. Transferred congenic, selfreactive CD4+ T cell clones rescue IgM-producing, scid-derived B cells.

Young severe combined immunodeficiency (scid) mice completely lack immunocompetent lymphocytes. Limiting numbers of purified CD4+ T cells from allotype-congenic BALB/c (Igha) donor mice were transplanted into 3-week-old scid (Ighb) recipient mice. Splenic CD4+ T cells were recovered from transplanted scid mice 10-12 weeks post-transfer and established as T cell lines in culture. These T cell populations proliferated in vitro in response to syngeneic stimulator cells. T cell clones derived in vitro from these T cell lines displayed selfreactive recognition specificity: these CD4+ T cells proliferated in vitro in response to syngeneic/congeneic but not allogeneic stimulator cells. Cloned selfreactive T cells retransplanted into young scid recipients were engrafted into spleens of secondary recipients, did not induce autoimmune disease but stimulated development of scid-derived (Ighb), IgM-producing B cells (B cell leakiness).

Animals↗
PubMed1991

CD3+ T cells in severe combined immunodeficiency (SCID) mice. V. Allogeneic T cells engrafted into SCID mice do not induce graft-versus-host disease in spite of the absence of host veto and natural suppressor cell activity.

Intravenous injection of 10(6) to 10(7) non-fractionated spleen cells (SC) from C57BL/6 (B6, H-2b) mice into completely allogeneic, immunodeficient H-2d severe combined immuno deficiency (scid) mice leads to engraftment of allogeneic donor T cells. Mice analysed in the tenth week posttransfer had engrafted donor-type CD4+ and CD8+ T cells in the spleens but showed no clinical evidence of graft-versus-host disease (GVHD). Transfer of allogeneic T cells engrafted in scid recipients did not induce GVHD upon i.v. injection into secondary scid recipients and lead in most recipients to engraftment of a pure CD4+ T-cell population. Experiments were carried out to investigate the reason(s) for the lack of GVHD in recipient scid mice, i.e. the presence of allotolerance in the engrafted donor T cells. Scid spleen cells (SC) efficiently stimulated alloreactive responses of B6 T cells: scid SC stimulated H-2d-specific cytotoxic responses in a B6 anti-scid mixed lymphocyte culture in vitro, and scid SC injected i.v. into B6 mice efficiently primed splenic cytotoxic lymphocyte precursors against H-2d alloantigens. Moreover when assayed in vitro, no veto activity or natural suppressor activity was detectable in scid SC. These data demonstrate that tolerizing mechanisms currently believed to operate in vivo can not explain the fact that allogeneic T cells injected i.v. into immunodeficient scid mice become tolerized against host-type alloantigens. Our results are discussed in the light of clinical experience of allogeneic T-cell transfer in scid infants.

Animals↗
PubMed1991

Selective reconstitution of T lymphocyte subsets in scid mice.

The 'empty' splenic T-cell compartment of young scid mice was partially and selectively reconstituted by low numbers of adoptively transferred congenic (C.B-17, BALB/c) or semi-allogeneic (dm2), but not completely allogeneic (C57BL/6) CD4+ T cells from adult donor mice. Under the same experimental conditions, the CD8+ T-cell compartment of young scid mice could only be reconstituted in scid mice continuously substituted with IL-2. CD4+ T cells engrafted in the spleen of H-2d scid recipients displayed specific, anti-Ad-associated proliferative reactivity. (Semi)allogeneic CD8+ T cells engrafted in spleens of scid recipients showed no anti-host H-2 class I-associated cytotoxic reactivity, i.e., this T-cell compartment was specifically tolerized against host (allo)antigens. Reconstitution of lymph nodes was not observed following transplantation of purified CD4+ or CD8+ T cells. Antigen-specific, cellular immune responses could not be induced in scid mice selectively reconstituted with either CD4+ T cells or CD8+ T cells. Partial reconstitution of the CD4+ T-cell compartment was always associated with the appearance of scid-derived, 'endogenous' IgM-producing B cells. Serum antibodies of scid mice with this T cell-induced B-cell leakiness stained an immature subset of congenic thymocytes.

Animals↗
PubMed1990

Characterization of cells with T cell markers in athymic nude bone marrow and of their in vitro-derived clonal progeny. Comparison with euthymic bone marrow.

Evidence is presented that early steps in T cell differentiation, to the point of TCR expression, can occur in athymic nude bone marrow (BM) and perhaps also in normal BM. Low density nylon wool nonadherent BM from 5- to 8-wk-old nude and normal mice was found to contain dim CD3+ CD4- CD8- cells. This subpopulation was not found in scid/scid low density nylon wool nonadherent BM but was present at similar frequency in both nude and normal BM and expressed comparable fluorescence intensity. With the use of additional markers, a difference between nude and normal dim CD3+ CD4- CD8- cells was found: the ratio of J11d+/J11d- cells was at least threefold higher in nude than in normal BM. Whereas 60% of nude J11d+ CD3+ cells expressed a TCR-alpha/beta and 40% expressed a gamma/delta TCR, almost no TCR-gamma/delta expressing cells were detected in dim CD3+ cells from normal mice in either the J11d+ or J11d- subpopulation. The majority of those cells express a TCR-alpha/beta. We established that nude dim CD3+ CD4- CD8- cells were distinguishable from mature recirculating T cells and developed an in vitro clonal culture system that supported the growth of these cells. When these cells were cocultured in vitro in a limiting dilution system with inactivated anti-CD3 mAb-secreting cells, in the presence of rIL-2 and rIL-4, one in 5 x 10(2) to 1 x 10(3) cells were growth inducible. Most progeny cells expressed a CD3+ CD8+ CD4- surface phenotype. Cultures displayed nonspecific cytolytic reactivity. With the use of a sensitive dot-blot technique, transcripts coding for C alpha, C beta, VC gamma, and C delta TCR genes were detected in these cultures. Rehybridization experiments demonstrated mutually exclusive expression of either C alpha or C delta mRNA transcripts in the majority of clones selected for a high probability of clonality. Most (greater than 95%) of T cell clones derived from normal BM expressed the C alpha transcript with approximately 5% expressing the C delta transcript. In contrast, about 40% of T cell clones derived from nude BM expressed the C delta transcript whereas the remaining 60% expressed 10-fold lower C alpha message than normal BM-derived clones.

Animals↗
PubMed1989

Diversity and structure of human T cell receptor delta chain genes in peripheral blood gamma/delta-bearing T lymphocytes.

We have investigated the diversity and repertoire of human TCR delta chain variable gene segments in the human peripheral blood CD4- CD8- (double-negative) population, using rearrangement and expression studies and sequence analyses. 20 TCR delta DNA clones were derived from the RNA of bulk-cultured double-negative T cells and their nucleotide sequences determined. These clones can be classified into six different V delta subfamilies. The distribution, however, was uneven in these cells, with 16 of 20 being derived from the V delta 1 (9) and V delta 2 (7) subfamilies. The remaining subfamilies, V delta 3, V delta 4, V delta 5, and V delta 6, were only represented by one clone each. The majority of these subfamilies seem to consist of a single member, in contrast with the closely linked V alpha subfamilies, which, in most cases, consist of multiple members. Our findings suggest that only a limited number of V delta genes are used in human peripheral blood double-negative T cells and that two major V delta subfamilies (V delta 1 and V delta 2) are used more frequently. Sequence comparison of our cDNA clones to V alpha clones indicates that there is no overlap in usage of V alpha and V delta gene segments, except for the V delta 4 (V alpha 6) subfamily. Comparison of the different V delta sequences suggests that the majority of the sequence diversity is concentrated in the junctions between V, D, and J segments and results from extensive N region diversity.

Antigens, Differentiation, T-Lymphocyte↗
PubMed1988

Sequence and organization of the human T cell delta chain gene.

A novel human T cell receptor (TcR) gene, located 85 kbp upstream to the C alpha coding regions, was isolated using human genomic clones to identify cDNA homologous to messages encoded by this region. The deduced protein sequence of this gene is highly homologous to that of the newly identified constant region found in the murine TcR alpha chain locus. This gene undergoes rearrangements and is expressed at the RNA level in human thymocytes, peripheral T cells and several leukemic T cell lines which have been shown to express the surface gamma-delta heterodimer, suggesting that this gene encodes the human T cell delta chain.

Amino Acid Sequence↗
PubMed1988

Development of autoreactive L3T4+ T cells from double-negative (L3T4-/Ly-2-) Thy-1+ spleen cells of normal mice.

Thy-1+/L3T4-/Ly-2- spleen cells were purified from normal C57BL/6 (B6) and C,B-17 mice. Cells within this subset expressed the T cell receptor (TcR) for antigen: the majority of cells in this subset were CD3+; a fraction of the cells was stained with the monoclonal antibody (mAb) F23.1; and the TcR molecule was immunoprecipitable with mAb F23.1 from cells within this subset. In limiting dilution analyses, about 1/30 cells within this subset were growth inducible in vitro by stimulation with phorbol myristate acetate (PMA) plus ionomycin; conditioned media containing interleukin (IL) 1, IL2, IL3 or IL4 activity neither triggered nor promoted in vitro growth of these cells. The in vitro generated T cells displayed the Thy-1+/L3T4+/Ly-2- surface phenotype and were self-reactive, i.e., proliferated preferentially in response to syngeneic stimulator cells, and secreted IL2 and IL3 only in response to syngeneic but not allogeneic stimulator cells. The proliferative response of these cells to syngeneic stimulator cells was blocked by anti-self Ia mAb. This autoreactive helper T cell subset was not inducible in purified Thy-1+ spleen cell subsets from athymic nude mice or scid mice. Autoreactive helper T cells did not express detectable levels of the IL2 receptor (IL2R), and their proliferative response was not blocked by anti-IL2R mAb. From PMA plus ionomycin-stimulated double-negative Thy-1+ spleen cells, 14 T cell clones were established in long-term culture which displayed the CD3+CD4+CD8- surface phenotype and were self-reactive.

Animals↗
PubMed1988

Sequence and organization of the diversity, joining, and constant region genes of the human T-cell delta-chain locus.

In this paper we describe the genomic organization and sequence of the human T-cell receptor delta-chain diversity, joining, and constant genes. There is one delta-chain constant region gene (C delta) located approximately equal to 85 kilobases (kb) upstream of the alpha-chain constant region. The delta-chain constant region consists of four exons, whose organization is very similar to that of the C alpha exons, suggesting that C alpha and C delta may have arisen from a gene duplication event. The first exon encodes most of the extracellular constant domain, the second encodes a hinge-like region, and the third encodes the entire transmembrane segment and intracytoplasmic portion, whereas the last exon contains exclusively 3' untranslated sequences. Three joining segments, J delta 1, J delta 2, and J delta 3, are found approximately equal to 12, approximately equal to 5.7, and approximately equal to 3.4 kb upstream of the first exon of C delta. Two functional diversity gene segments, D delta 1 and D delta 2, which can be productively translated in all three reading frames, are found 1 and 9.6 kb upstream of J delta 1. The presence of two D delta with such potential for diversity may offset the limited repertoire of the J delta and V delta genes. The spacer distribution in the recombinational signals flanking D delta and J delta segments allows recombination with V alpha gene segments; however, examination of delta-chain messages does not indicate that this is the case, suggesting that the delta chain uses unique variable gene segments and raising the question as to the reasons for this phenomenon.

Base Sequence↗