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In vitro induction of CD8 expression on thymic pre-T cells. II. Characterization of CD3-CD4-CD8 alpha + cells generated in vitro by culturing CD25+CD3-CD4-CD8- thymocytes with T cell growth factor-beta and tumor necrosis factor-alpha.
J Immunol. 1992 Jul 01; 149(1):71-6.JI

Abstract

We previously reported that CD25 (IL-2R p55)-positive CD3-CD4-CD8- murine thymocytes can be induced to express CD8 alpha (Lyt-2) by transforming growth factor-beta plus TNF-alpha in the presence of IL-7 (which is necessary to maintain the viability and differentiation capacity of CD25+CD3-CD4-CD8- thymocytes in vitro). The majority of cells recovered after 2 to 3 days from these cultures expressed CD8 alpha (but not CD3 or CD4). In this study, we have characterized these in vitro generated CD3-CD4-CD8 alpha + thymocytes and compared them with normal CD3-CD4-CD8+ thymocytes. Unlike normal CD3-CD4-CD8+ thymocytes that express CD8 alpha and CD8 beta (Lyt-3-chain) simultaneously, only a fraction of in vitro generated CD3-CD4-CD8 alpha + cells expressed CD8 beta. However, along with the induction of CD8 alpha and CD8 beta expression, the expression of other T cell differentiation markers (including CD2, CD25, and CD44) also changed in a manner corresponding to physiologic differentiation. Cell-surface phenotyping suggests that CD8 alpha + beta - cells are less mature than CD8 alpha + beta + cells. These in vitro generated CD3-CD4-CD8 alpha + thymocytes expanded and differentiated into the CD4+CD8+ stage as well as mature (CD3+) single positive (CD4+CD8-) and CD4-CD8+) stages in fetal thymus organ culture that had been depleted of lymphoid cells by treatment with 2-deoxyguanosine. The latter observation indicates that these in vitro generated CD3-CD4-CD8 alpha + thymocytes are responsive to other differentiation-inducing signals (including those that induce CD4) that exist in fetal thymus organ culture. These results suggest that in vitro generated CD3-CD4-CD8 alpha + thymocytes represent intermediate differentiation stages between CD25+CD3-CD4-CD8- and CD3-CD4-CD8+ cells found in normal thymus.

Authors+Show Affiliations

DNAX Research Institute of Molecular and Cellular Biology, Palo Alto, CA 94304.No affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

1607663

Citation

Suda, T, and A Zlotnik. "In Vitro Induction of CD8 Expression On Thymic pre-T Cells. II. Characterization of CD3-CD4-CD8 Alpha + Cells Generated in Vitro By Culturing CD25+CD3-CD4-CD8- Thymocytes With T Cell Growth Factor-beta and Tumor Necrosis Factor-alpha." Journal of Immunology (Baltimore, Md. : 1950), vol. 149, no. 1, 1992, pp. 71-6.
Suda T, Zlotnik A. In vitro induction of CD8 expression on thymic pre-T cells. II. Characterization of CD3-CD4-CD8 alpha + cells generated in vitro by culturing CD25+CD3-CD4-CD8- thymocytes with T cell growth factor-beta and tumor necrosis factor-alpha. J Immunol. 1992;149(1):71-6.
Suda, T., & Zlotnik, A. (1992). In vitro induction of CD8 expression on thymic pre-T cells. II. Characterization of CD3-CD4-CD8 alpha + cells generated in vitro by culturing CD25+CD3-CD4-CD8- thymocytes with T cell growth factor-beta and tumor necrosis factor-alpha. Journal of Immunology (Baltimore, Md. : 1950), 149(1), 71-6.
Suda T, Zlotnik A. In Vitro Induction of CD8 Expression On Thymic pre-T Cells. II. Characterization of CD3-CD4-CD8 Alpha + Cells Generated in Vitro By Culturing CD25+CD3-CD4-CD8- Thymocytes With T Cell Growth Factor-beta and Tumor Necrosis Factor-alpha. J Immunol. 1992 Jul 1;149(1):71-6. PubMed PMID: 1607663.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - In vitro induction of CD8 expression on thymic pre-T cells. II. Characterization of CD3-CD4-CD8 alpha + cells generated in vitro by culturing CD25+CD3-CD4-CD8- thymocytes with T cell growth factor-beta and tumor necrosis factor-alpha. AU - Suda,T, AU - Zlotnik,A, PY - 1992/7/1/pubmed PY - 1992/7/1/medline PY - 1992/7/1/entrez SP - 71 EP - 6 JF - Journal of immunology (Baltimore, Md. : 1950) JO - J Immunol VL - 149 IS - 1 N2 - We previously reported that CD25 (IL-2R p55)-positive CD3-CD4-CD8- murine thymocytes can be induced to express CD8 alpha (Lyt-2) by transforming growth factor-beta plus TNF-alpha in the presence of IL-7 (which is necessary to maintain the viability and differentiation capacity of CD25+CD3-CD4-CD8- thymocytes in vitro). The majority of cells recovered after 2 to 3 days from these cultures expressed CD8 alpha (but not CD3 or CD4). In this study, we have characterized these in vitro generated CD3-CD4-CD8 alpha + thymocytes and compared them with normal CD3-CD4-CD8+ thymocytes. Unlike normal CD3-CD4-CD8+ thymocytes that express CD8 alpha and CD8 beta (Lyt-3-chain) simultaneously, only a fraction of in vitro generated CD3-CD4-CD8 alpha + cells expressed CD8 beta. However, along with the induction of CD8 alpha and CD8 beta expression, the expression of other T cell differentiation markers (including CD2, CD25, and CD44) also changed in a manner corresponding to physiologic differentiation. Cell-surface phenotyping suggests that CD8 alpha + beta - cells are less mature than CD8 alpha + beta + cells. These in vitro generated CD3-CD4-CD8 alpha + thymocytes expanded and differentiated into the CD4+CD8+ stage as well as mature (CD3+) single positive (CD4+CD8-) and CD4-CD8+) stages in fetal thymus organ culture that had been depleted of lymphoid cells by treatment with 2-deoxyguanosine. The latter observation indicates that these in vitro generated CD3-CD4-CD8 alpha + thymocytes are responsive to other differentiation-inducing signals (including those that induce CD4) that exist in fetal thymus organ culture. These results suggest that in vitro generated CD3-CD4-CD8 alpha + thymocytes represent intermediate differentiation stages between CD25+CD3-CD4-CD8- and CD3-CD4-CD8+ cells found in normal thymus. SN - 0022-1767 UR - https://www.unboundmedicine.com/medline/citation/1607663/In_vitro_induction_of_CD8_expression_on_thymic_pre_T_cells__II__Characterization_of_CD3_CD4_CD8_alpha_+_cells_generated_in_vitro_by_culturing_CD25+CD3_CD4_CD8__thymocytes_with_T_cell_growth_factor_beta_and_tumor_necrosis_factor_alpha_ L2 - https://www.jimmunol.org/lookup/pmidlookup?view=long&pmid=1607663 DB - PRIME DP - Unbound Medicine ER -