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1.
Sci Rep ; 5: 9219, 2015 Mar 18.
Article in English | MEDLINE | ID: mdl-25783199

ABSTRACT

The T cell specific adapter protein (TSAd) is expressed in activated T cells and NK cells. While TSAd is beginning to emerge as a critical regulator of Lck and Itk activity in T cells, its role in NK cells has not yet been explored. Here we have examined susceptibility to virus infections in a murine model using various viral infection models. We report that TSAd-deficient mice display reduced clearance of murine cytomegalovirus (MCMV) that lack the viral MHC class I homologue m157, which is critical for Ly49H-mediated NK cell recognition of infected cells. In this infection model, NK cells contribute in the early stages of the disease, whereas CD8+ T cells are critical for viral clearance. We found that mice infected with MCMV Δm157 displayed reduced viral clearance in the spleen as well as reduced proliferation in spleen NK cells and CD8+ T cells in the absence of TSAd. Though no other immunophenotype was detected in the infection models tested, these data suggests that in the absence of the Ly49H ligand activation, NK cell and CD8+ T cell responses may be compromised in TSAd-deficient mice.


Subject(s)
Adaptor Proteins, Signal Transducing/metabolism , Muromegalovirus/genetics , Viral Proteins/genetics , Adaptor Proteins, Signal Transducing/genetics , Animals , CD11b Antigen/metabolism , CD8-Positive T-Lymphocytes/cytology , CD8-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/metabolism , Cell Line , Cell Proliferation , Cytomegalovirus Infections/immunology , Cytomegalovirus Infections/veterinary , Cytomegalovirus Infections/virology , Disease Models, Animal , Flow Cytometry , Genotype , Humans , Immunophenotyping , Killer Cells, Natural/cytology , Killer Cells, Natural/immunology , Killer Cells, Natural/metabolism , Mice , Mice, Inbred C57BL , Mice, Knockout , Muromegalovirus/physiology , Mutation , Spleen/cytology , Spleen/immunology , Spleen/virology , Tumor Necrosis Factor Receptor Superfamily, Member 7/metabolism , Viral Load , Virus Replication
2.
Scand J Immunol ; 80(3): 169-79, 2014 Sep.
Article in English | MEDLINE | ID: mdl-24910151

ABSTRACT

T cell-specific adapter protein (TSAd) encoded by the SH2D2A gene is expressed in activated T cells, NK cells and endothelial cells, but its tissue expression has not yet been mapped. Here, we have defined the specificity of two commercially available anti-TSAd monoclonal reagents using peptide arrays. We found them to bind separate epitopes in the C-terminal part of TSAd. We then used immunohistochemistry to examine TSAd expression in various human lymphoid and non-lymphoid tissues. Immunostaining of adjacent tissue sections revealed that a substantial fraction of CD3-positive cells in normal lymphoid and non-lymphoid tissues expressed TSAd. In particular, essentially all intra-epithelial T cells appeared to coexpress TSAd. In addition, TSAd expression was observed in endothelial cells of dermal microvessels, while it was not detected in endothelial cells of the other tested tissues. This work provides insight into the expression pattern of TSAd in various healthy human tissues.


Subject(s)
Adaptor Proteins, Signal Transducing/biosynthesis , Endothelial Cells/metabolism , Lymphoid Tissue/metabolism , T-Lymphocytes/metabolism , Adaptor Proteins, Signal Transducing/immunology , Amino Acid Sequence , Antibodies, Monoclonal/immunology , Antibodies, Monoclonal/metabolism , CD3 Complex/immunology , CD3 Complex/metabolism , Dermis/blood supply , Endothelial Cells/immunology , Epithelium/immunology , Epithelium/metabolism , Epitopes/immunology , Epitopes/metabolism , Humans , Immunohistochemistry/methods , Lymphoid Tissue/immunology , Microvessels/cytology , Microvessels/immunology , Microvessels/metabolism , Molecular Sequence Data , Peptides/immunology , Peptides/metabolism , T-Lymphocytes/immunology
3.
J Anim Sci ; 90(2): 481-8, 2012 Feb.
Article in English | MEDLINE | ID: mdl-21984713

ABSTRACT

Regulations for the disposal of genetically engineered animals are strict due to concern for their inappropriate introduction into the food chain, and of the possible public health and environmental impacts of these organisms. Nontransgenic animals that give birth to transgenic offspring are treated as if they are transgenic due to concern of fetal cells crossing the placental barrier and residing in the mother (fetal-maternal microchimerism). Determining whether or not fetal-fetal or fetal-maternal transfer of DNA or cells occurs during caprine gestation is critical to effectively protect the public without culling animals that pose no risk. Additionally, fetal-maternal transfer, should it exist in the goat, could contraindicate the rebreeding of nontransgenic dams due to the possible transfer of fetal cells from 1 pregnancy to the fetus of subsequent pregnancies. Fetal-maternal transfer in Capra hircus has not been reported in the literature, although it has been reported in another ruminant, Bos taurus. We examined blood from nontransgenic dams that carried transgenic offspring using a PCR method sensitive enough to detect the presence of a spider silk transgene to a 1:100,000 dilution. At this sensitivity, we did not detect the occurrence of fetal-maternal transfer in 5 nontransgenic dams. Likewise, fetal-fetal transfer was not observed from a transgenic to a nontransgenic twin in utero. To test tissue-specific expression of the silk transgene, proteins purified from standard necropsy tissue from a lactating transgenic dam were examined by Western blot analysis. Silk protein expression was only observed in mammary tissue consistent with the tissue specificity of the ß-casein promoter used in the transgenic construct. We report evidence collected from a limited caprine breeding pool against transfer of transgenes in utero from fetus to dam and fetus to fetus. In addition, we show evidence that the ß-casein promoter in our expression construct is not expressed ectopically as previously suggested. These results suggest that transgene transfer in utero does not occur, but further study is warranted with a larger sample group to confirm these results.


Subject(s)
Caseins/genetics , Chimerism , Fibroins/genetics , Goats/genetics , Animals , Animals, Genetically Modified , Animals, Newborn , Blotting, Western , DNA/chemistry , DNA/genetics , Female , Fibroins/analysis , Male , Polymerase Chain Reaction/veterinary , Pregnancy
5.
Eur J Immunol ; 30(12): 3355-62, 2000 Dec.
Article in English | MEDLINE | ID: mdl-11093152

ABSTRACT

Here we report the generation of monoclonal antibodies (mAb) LOV3 and LOV8 to a 110-130-kDa membrane glycoprotein expressed by rat NK cells. This NK surface molecule was identified by eucaryotic expression cloning as the structural orthologue of the phagocytosis-stimulating receptor for complement factor C1q and mannose-binding lectin on human macrophages, C1qRp. Rat C1qRp is a monomeric type I integral membrane protein consisting of 643 amino acids with an N-terminal lectin-like domain, five epidermal growth factor-like domains, a transmembrane domain and a 45-residue cytoplasmic domain. It is encoded by a single gene on rat chromosome 3q41-q42 and is 67% and 87.5% identical at the amino acid level to human and mouse C1qRp, respectively. Rat C1qRp is expressed by resting and by activated NK cells, on subpopulations of NKR-P1(+) T cells (NK/T cells), dendritic cells, macrophages and granulocytes, but not by B cells or NKR-P1(-) T cells. Expression of this innate immune receptor is therefore not restricted to hematopoietic cells of the myeloid lineage, but is also expressed on subsets of cells of lymphoid origin. The mAb did not affect the cytotoxic function of NK cells, and C1qRp on NK cells may have functions not related to NK killing.


Subject(s)
Hyaluronan Receptors , Killer Cells, Natural/chemistry , Membrane Glycoproteins , Receptors, Cell Surface/genetics , Receptors, Complement/genetics , Amino Acid Sequence , Animals , Carrier Proteins , Chromosome Mapping , Cloning, Molecular , Humans , Mice , Mice, Inbred BALB C , Mitochondrial Proteins , Molecular Sequence Data , Rats , Receptors, Complement/analysis , Receptors, Complement/chemistry
7.
Eur J Immunol ; 29(6): 2000-6, 1999 06.
Article in English | MEDLINE | ID: mdl-10382763

ABSTRACT

Several receptors expressed by subsets of leukocytes and with sequence homology to the killer cell inhibitory receptors have recently been identified both in man and mouse. Here we describe a rat cDNA that encodes a novel receptor of this group, designated neutrophil immunoglobulin-like receptor-1 (NILR-1). The predicted 58.7-kDa mature NILR-1 protein is a type I integral membrane protein, with three C2-type immunoglobulin superfamily domains, a transmembrane region devoid of charged amino acids, and a cytoplasmic tail containing four immunoreceptor tyrosine-based inhibition motif-like regions. NILR-1 shows greatest sequence homology to the mouse paired immunoglobulin-like receptor-B and members of the human leukocyte immunoglobulin-like receptor/immunoglobulin-like transcript group of receptors. As shown by Northern blot analysis, NILR-1 was transcribed by neutrophilic granulocytes. Although weaker transcription was found with a macrophage cell line, no signal was detected with peritoneal macrophage or spleen RNA. Linkage analysis localized Nilr1 to chromosome 1, closely linked to a locus encoding a rat NKp46 orthologue. The two loci define a rat leukocyte receptor gene complex, in a region syntenic to human chromosome 19q13.4 and the proximal part of mouse chromosome 7, that harbors the human and mouse leukocyte receptor gene complexes.


Subject(s)
Immunoglobulins/genetics , Neutrophils/immunology , Receptors, Immunologic/genetics , Amino Acid Sequence , Animals , Chromosome Mapping , Cloning, Molecular , DNA, Complementary/genetics , Gene Expression , Humans , Male , Mice , Molecular Sequence Data , RNA, Messenger/genetics , RNA, Messenger/metabolism , Rats , Rats, Inbred Strains , Receptors, Immunologic/chemistry , Sequence Homology, Amino Acid , Species Specificity , Tissue Distribution
8.
Eur J Immunol ; 28(6): 1839-46, 1998 Jun.
Article in English | MEDLINE | ID: mdl-9645365

ABSTRACT

Three classes of multigene family-encoded receptors enable NK cells to discriminate between polymorphic MHC class I molecules: Ly-49 homodimers, CD94/NKG2 heterodimers and the killer cell inhibitory receptors (KIR). Of these, CD94/NKG2 has been characterized in both rodents and humans. In contrast, Ly-49 family members have hitherto been found only in rodents, and KIR molecules only in the human. In this report, we describe a human cDNA, termed Ly-49L, that constitutes the first human member of the Ly-49 multi-gene family. Compared with rodent Ly-49 molecules, the Ly-49L sequence contains a premature stop codon and predicts a truncated protein that lacks the distal part of a C-terminal lectin domain. Evidence is presented that the premature stop codon results from incomplete excision of the intron between the first two lectin domain exons. Splice variants predicting a full-size Ly-49L protein were not detected. As demonstrated by Northern blot analysis, Ly-49L was transcribed by IL-2-activated NK cells, but not by freshly isolated B or T cells. PCR screening of a 22-clone yeast artificial chromosome contig localized the LY49L locus to the human NK gene complex on chromosome 12p12-p13. Southern blot analysis of genomic DNA showed a simple pattern with a full-length Ly-49L probe at low stringency hybridization conditions, suggesting that Ly-49L may be the only human member of the Ly-49 multigene family.


Subject(s)
Antigens, Ly/genetics , Multigene Family , Alternative Splicing , Amino Acid Sequence , Animals , Antigens, Ly/classification , Base Sequence , Chromosome Mapping , DNA, Complementary , Exons , Humans , Killer Cells, Natural/metabolism , Mice , Molecular Sequence Data , Rats , Sequence Analysis, DNA , Sequence Homology, Amino Acid , Transcription, Genetic
9.
Int Immunol ; 10(4): 379-85, 1998 Apr.
Article in English | MEDLINE | ID: mdl-9620593

ABSTRACT

The gene for a rat NK lectin-like receptor (NKLLR), named NKR-P2, has been cloned and characterized. Sequence analysis shows that it represents the orthologue of human NKG2D and that the two molecules form a distinct NKLLR family, no more related to NKG2A/B, -C or -E than to other NKLLR families. Nkrp2 is a single-copy gene containing seven introns, mapping to the rat NK gene complex. Rat NKR-P2 differs from the human orthologue in that its cytoplasmic tail contains 13 additional amino acids, encoded by a separate exon. Splice variants lacking this exon were not detected in T cells or NK cells. NKR-P2 is strongly expressed by NK cells. In contrast to other NKLLR, it is also strongly expressed by resting thoracic duct CD4+ and CD8+ T cells, but not by thymocytes or other hemopoietic cells.


Subject(s)
CD4-Positive T-Lymphocytes/ultrastructure , CD8-Positive T-Lymphocytes/ultrastructure , Killer Cells, Natural/ultrastructure , Lectins, C-Type , Lectins/metabolism , Receptors, Immunologic/biosynthesis , Receptors, Immunologic/genetics , Alternative Splicing , Amino Acid Sequence , Animals , Chromosome Mapping , Cloning, Molecular , DNA, Complementary/genetics , Exons , Introns , Molecular Sequence Data , NK Cell Lectin-Like Receptor Subfamily K , Rats , Rats, Inbred F344 , Sequence Homology, Amino Acid
10.
Eur J Immunol ; 28(2): 444-50, 1998 Feb.
Article in English | MEDLINE | ID: mdl-9521051

ABSTRACT

Two different lectin-like receptors for MHC class I molecules have so far been identified on natural killer (NK) cells, the Ly-49 homodimeric receptors in mice and the NKG2/CD94 heterodimeric receptors in humans. The recent identification of a rat CD94 orthologue implied that NK cell receptors equivalent to NKG2/CD94 also exist in rodents. Here we describe the cDNA cloning of two rat genes homologous to members of the human NKG2 multigene family. The deduced rat NKG2A protein contains a cytoplasmic immunoreceptor tyrosine-based inhibition motif (ITIM), whereas the cytoplasmic tail of rat NKG2C lacks ITIM. The genes map to the rat NK gene complex and are selectively expressed by NK cells. The expression is strain dependent, with high expression in DA and low in PVG NK cells, correlating with the expression of rat CD94. Ly-49 genes have previously been identified in the rat, and the existence of rat NKG2 genes in addition to a CD94 orthologue suggests that NK cell populations utilize different C-type lectin receptors for MHC class I molecules in parallel.


Subject(s)
Antigens, CD/genetics , Killer Cells, Natural/metabolism , Lectins, C-Type , Membrane Glycoproteins/genetics , Sequence Homology, Amino Acid , Sequence Homology, Nucleic Acid , Amino Acid Sequence , Animals , Antigens, CD/biosynthesis , Antigens, Surface/genetics , DNA, Complementary/isolation & purification , Humans , Membrane Glycoproteins/biosynthesis , Molecular Sequence Data , NK Cell Lectin-Like Receptor Subfamily B , NK Cell Lectin-Like Receptor Subfamily C , NK Cell Lectin-Like Receptor Subfamily D , Polymorphism, Restriction Fragment Length , Rats , Receptors, Immunologic/genetics , Receptors, Natural Killer Cell , Sequence Analysis, DNA
11.
Eur J Immunol ; 27(8): 2080-6, 1997 Aug.
Article in English | MEDLINE | ID: mdl-9295048

ABSTRACT

Three classes of major histocompatibility (MHC) class I binding receptors on natural killer (NK) cells have so far been described: CD94/NKG2 heterodimeric receptors and killer cell inhibitory receptors in the human, and Ly-49 homodimers in rodents. CD94, NKG2 and Ly-49 belong to the C-type lectin superfamily. As yet, CD94 and NKG2 molecules have not been detected in rodents or Ly-49 in humans. It has therefore been proposed that the two receptors represent functional equivalents in these species. The present study describes the cDNA cloning of a novel rat gene encoding a protein of 179 amino acids, 54.2% identical to human CD94. The single-copy Cd94 gene is localized to the rat NK gene complex (NKC), within 50 kb from Nkrp2, between the Nkrp1 and Ly49 gene clusters. By Northern blot analysis, we showed that rat CD94 is selectively expressed by NK cells and a small subset of T cells, similar to the human orthologue. This expression is strain dependent, with high expression in DA NK cells and low in PVG NK cells. Evidence is presented that this difference is not due to receptor repertoire shaping by MHC-encoded ligands, but is controlled by genetic elements residing within the NKC. The identification of a rat CD94 orthologue suggests that NK cell populations utilize two different C-type lectin receptors for MHC class I molecules in parallel.


Subject(s)
Antigens, CD/genetics , Killer Cells, Natural/immunology , Lectins, C-Type , Membrane Glycoproteins/genetics , Amino Acid Sequence , Animals , Chromosome Mapping , Cloning, Molecular , DNA, Complementary/genetics , Female , Gene Expression , Genetic Linkage , Humans , Male , Molecular Sequence Data , NK Cell Lectin-Like Receptor Subfamily D , Rats , Rats, Inbred Strains , Sequence Homology, Amino Acid , Species Specificity
12.
J Exp Med ; 183(5): 2197-207, 1996 May 01.
Article in English | MEDLINE | ID: mdl-8642329

ABSTRACT

Natural Killer (NK) cells can recognize and kill MHC-incompatible normal bone marrow-derived cells. Presently characterized MHC-binding receptors on NK cells, including the Ly-49 family in the mouse, transmit inhibitory signals upon binding to cognate class I MHC ligands. Here we study in vivo NK-mediated lysis of normal allogeneic lymphocytes in crosses between alloreactivity-competent PVG rats and alloreactivity-deficient DA rats. NK cells from both strains are able to lyse standard tumor targets. We identify an autosomal dominant locus, Nka, that controls NK-mediated alloreactivity. Individuals carrying the dominant PVG allele in single dose were fully competent in eliminating allogeneic target cells, suggesting that Nka encodes or regulates a gene product inducing or activating alloreactivity. By linkage analysis and pulsed field gel electrophoresis, a natural killer gene complex (NKC) on rat chromosome 4 is described that contains the rat NKR-P1 and Ly-49 multigene families plus a rat NKG2D homologue. Nka maps within the NKC, together with the most telomeric Ly-49 family members, but separate from NKG2D and the NKR-P1 family. The Nka-encoded response, moreover, correlates with the expression of transcripts for Ly-49 receptors in NK cell populations, as Northern blot analysis demonstrated low expression of Ly-49 genes in DA NK cells, in contrast to high expression in alloreactivity-competent PVG, (DA X PVG)F1, and PVG.1AVI NK cells. The low Ly-49 expression in DA is not induced by MHC haplotype, as demonstrated by high expression of Ly-49 in the DA MHC-congenic PVG.1AVI strain. Finally, we have cloned and characterized the first four members of the rat Ly-49 gene family. Their cytoplasmic domains demonstrate substantial heterogeneity, consistent with the hypothesis that different Ly-49 family members may subserve different signaling functions.


Subject(s)
Antigens, Ly , Chromosome Mapping , Genes, Dominant , Killer Cells, Natural/immunology , Membrane Glycoproteins/genetics , Amino Acid Sequence , Animals , Antigens, Surface/immunology , Base Sequence , Consensus Sequence , Crosses, Genetic , DNA Primers , Exons , Histocompatibility Antigens Class I/immunology , Isoantigens/immunology , Lectins, C-Type , Major Histocompatibility Complex , Membrane Glycoproteins/biosynthesis , Mice , Molecular Sequence Data , Phylogeny , Polymerase Chain Reaction , Pseudogenes , Rats , Rats, Inbred F344 , Rats, Inbred Lew , Rats, Inbred Strains , Receptors, NK Cell Lectin-Like , Sequence Homology, Amino Acid
15.
Eur J Immunol ; 21(9): 2167-75, 1991 Sep.
Article in English | MEDLINE | ID: mdl-1716212

ABSTRACT

We have previously shown that large granular lymphocyte (LGL)-enriched cell populations have the capacity to spontaneously recognize and kill allogeneic small lymphocytes and bone marrow cells (BMC) in vitro in certain strain combinations of rats. Here, we have studied the alloreactivity of natural killer (NK) cells from PVG nude (RT1c) rats against a panel of major histocompatibility complex (MHC) incompatible hemic cells. Both lymphocytes and BMC from the AO (RT1u), DA (RT1a), BN (RT1n) as well as the MHC-congenic PVG-RT1u (RT1u) rat strains were efficiently killed in vitro, whereas cells from syngeneic PVG rats were spared. The structures recognized on lymphocytes and BMC were probably similar since the two cell populations inhibited each other in cross-competition experiments. A number of features aligned the alloreactive effector cells with NK cells and not T cells. (a) Only about 5% of the effector cells from nude spleens expressed the T cell antigens CD3, CD5 or T cell receptor (TcR) alpha/beta whereas greater than 50% of the cells expressed markers present on NK cells (CD2, CD8, OX52 and the rat NK cell-specific marker NKR-P1 recognized by the monoclonal antibody 3.2.3). (b) The alloreactive cells were granular since pretreatment of nude spleen cells with the lysosomotropic agent L-leucine methyl ester which eliminated LGL, simultaneously abolished the cytolysis of both allogeneic lymphocytes and YAC-1 tumor cells. (c) Nude spleen cells stimulated with human recombinant interleukin 2 for 1 week in vitro generated large granular proliferating cells which were CD3-, CD5-, TcR alpha/beta-, but greater than 95% 3.2.3+. These cells efficiently killed allogeneic hemic cells from the same rat strains as did freshly isolated effector cells. (d) The cytolysis of allogeneic hemic cells could effectively be inhibited with unlabelled NK-sensitive (YAC-1 and K-562), but not NK-resistant (Roser leukemia) tumor cells. Cross-competition studies showed that PVG nude NK cells discriminated between AO, BN and DA BMC, suggesting that different alloantigens were positively recognized by subsets of NK cells. The mode of inheritance of the allodeterminant specifically recognized on AO BMC was investigated in crosses and backcrosses between AO and BN or DA rats. A gene dosage effect was observed in that this determinant was expressed at a slightly reduced level in F1 hybrids.(ABSTRACT TRUNCATED AT 250 WORDS)


Subject(s)
Antibody-Dependent Cell Cytotoxicity , Bone Marrow/immunology , Killer Cells, Natural/immunology , Major Histocompatibility Complex/immunology , Animals , Cross Reactions , Crosses, Genetic , Cytotoxicity Tests, Immunologic , Dose-Response Relationship, Immunologic , Epitopes , Flow Cytometry , Haplotypes , Immunophenotyping , In Vitro Techniques , Interleukin-2/pharmacology , Leucine/analogs & derivatives , Leucine/pharmacology , Lymphocytes/immunology , Rats , Rats, Nude , Spleen/immunology
16.
Eur J Immunol ; 20(5): 1193-6, 1990 May.
Article in English | MEDLINE | ID: mdl-2162777

ABSTRACT

Rearrangement of both the beta and gamma chain T cell receptor (TcR) genes was detected in intestinal intraepithelial lymphocytes (IEL) from normal euthymic rats. Flow cytometric analyses showed that about 73% of the IEL were CD3+ (1F4) and that 67% were TcR alpha/beta+ (R73). About 5% of the IEL were found to be CD3+, TcR alpha/beta- in double-labeling experiments suggesting that a small fraction of IEL in the rat express the alternative TcR gamma/delta. More than 70% of the IEL were granular implying that many CD3+ IEL are granular. In IEL from athymic nude rats no rearrangement of either the TcR beta or gamma chain genes or surface expression of CD3 or TcR alpha/beta was detected despite the fact that about 95% of the cells were granular and morphologically similar to those in normal rats. Taken together our data suggest that the majority of IEL in the rat express the conventional TcR alpha/beta and that TcR-bearing cells in the gut epithelium are thymus dependent.


Subject(s)
Intestinal Mucosa/immunology , Receptors, Antigen, T-Cell/genetics , T-Lymphocytes/analysis , Thymus Gland/physiology , Animals , Gene Rearrangement, alpha-Chain T-Cell Antigen Receptor/physiology , Gene Rearrangement, beta-Chain T-Cell Antigen Receptor/physiology , Phenotype , Rats , Rats, Nude , Receptors, Antigen, T-Cell, alpha-beta
17.
Int Immunol ; 2(5): 453-60, 1990.
Article in English | MEDLINE | ID: mdl-2150758

ABSTRACT

Lymphokine activated killer (LAK) cells from athymic nude rats, previously shown to selectively kill MHC-incompatible small lymphocytes in vitro, were examined for rearrangement and expression of genes for the T cell antigen receptors. Southern blots showed no rearrangement of the TCR beta-chain genes, and Northern blots showed transcription only of truncated 1.0 kb beta-chain messages, but not of full-length, 1.3 kb beta-chain mRNA. Transcription of the alpha-chain of the TCR could not be detected, and surface staining with the mAb R73 showed no expression of the rat TCR alpha/beta heterodimer. Transcripts hybridizing with a rat TCR C gamma probe were detected on Northern blots, but probes for all presently characterized mouse V gamma genes failed to hybridize to the same filters, indicating that the C gamma-containing transcripts probably were from non-rearrangement genes. CD3 delta- and epsilon-chain transcripts could not be detected by Northern blot analysis. Less than 2% of the cells stained with the anti-rat CD3 monoclonal antibody 1F4, and incubation with 1F4 had no effect on the alloreactivity of nude rat LAK cells. We have previously shown that immunoglobulin is not involved in the killing of the allogeneic lymphocytes. The most likely interpretation of these results is therefore that nude rat LAK cells express a novel receptor structure involved in allorecognition.


Subject(s)
Antigens, Differentiation, T-Lymphocyte/metabolism , Killer Cells, Lymphokine-Activated/immunology , Rats, Nude/immunology , Receptors, Antigen, T-Cell/metabolism , Animals , Antigens, Differentiation, T-Lymphocyte/genetics , CD3 Complex , Cytotoxicity, Immunologic , Gene Rearrangement, T-Lymphocyte , Rats , Rats, Nude/genetics , Receptors, Antigen, T-Cell/genetics , Transcription, Genetic
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