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Receptor specificity, functional characteristics, and cell-surface phenotype of a highly selected anti-I-Ab-specific, long-term T-cell line

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Abstract

A highly selected alloreactive T-cell line was developed by repeated restimulation of B10.D2/n lymph-node cells with irradiated C57BL/10Sn (BIO) spleen cells in long-term MLC for up to 2 1/2 years. Continuous growth of the line requires restimulation every 2 to 4 weeks with fresh H-2b stimulator cells. The line proliferates strongly against H-2b but not againstH-2 d,H-2 f,H-2 q,H-2 r, orH-2 s stimulators. Analysis of recombinant mouse strains showed that the proliferative response is directed against I-Ab but not Kb or Db determinants. During the growth period of the line, strong cross-reactivity with H-2p (B10.P) and weak cross-reactivity with H-2k strains (e.g., CBA/J and B10.BR) was observed. A clone with exquisite specificity for I-Ab, but with no cross-reactivity with H-2p or H-2k was isolated from the line; thus clonal heterogeneity of the line still exists despite the highly selective growth conditions. — The majority of T cells from the line or clone were shown to bind I-Ab but not Kb or Db determinants either spontaneously during restimulation with fresh B10 stimulator cells or via membrane vesicles expressing I-Ab determinants. No killing activity by the line in either specific or nonspecific cytolytic T-cell assays was observed nor was the T 145 glycoprotein, characteristic of killer T cells, detected.

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Abbreviations

B6:

C57BL/6J

B10:

C57BL/10Sn

Con A:

Concanavalin A

CTL:

cytotoxic T lymphocyte

FCS:

fetal calf serum

FDA:

fluorescein diacetate

FITC:

fluorescein isothiocyanate

Ia:

I-region-associated antigens

LPS:

lipopolysaccharide fromE. coli

Lyt:

T-lymphocyte-defined antigen

MLC:

mixed leukocyte culture

NP-40:

nonidet P-40

PAGE:

pofyacrylamide gel electrophoresis

PHA:

phytohemagglutinin fromPhaseolus vulgaris

PM:

plasma membrane

SDS:

sodium dodecyl sulfate

TCGF:

T-cell growth factor(s)

TdR:

thymidine

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Author notes

  1. Ammon B. Peck

    Present address: Department of Pathology, College of Medicine, University of Florida, 32610, Gainesville, Florida

  2. Zoltan A. Nagy

    Present address: Department of Immunogenetics, Max Planck Institute for Biology, 7400, Tübingen 1, Germany

Authors and Affiliations

  1. Cancer Research Laboratories, Botterell Hall, Department of Pathology, Queen's University, K7L 3N6, Kingston, Ontario, Canada

    Bruce E. Elliott (Research Scholar of the National Cancer Institute of Canada) & Zoltan A. Nagy

  2. Department of Immunology, University of Uppsala Biomedical Center, Box 582, S-751, Uppsala, Sweden

    Arthur Kimura, Ammon B. Peck & Hans Wigzell

Authors
  1. Bruce E. Elliott
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  2. Arthur Kimura
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  3. Ammon B. Peck
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  4. Zoltan A. Nagy
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  5. Hans Wigzell
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Elliott, B.E., Kimura, A., Peck, A.B. et al. Receptor specificity, functional characteristics, and cell-surface phenotype of a highly selected anti-I-Ab-specific, long-term T-cell line. Immunogenetics 12, 525–540 (1981). https://doi.org/10.1007/BF01561693

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  • DOI: https://doi.org/10.1007/BF01561693

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