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Novel genetic regulation of T helper 1 (Th1)/Th2 cytokine production and encephalitogenicity in inbred mouse strains.

Conboy IM, DeKruyff RH, Tate KM, Cao ZA, Moore TA, Umetsu DT, Jones PP - J. Exp. Med. (1997)

Bottom Line: A splenic APC reduced IFN-gamma and TNF-alpha production by established Th1 MBP-specific Ak-restricted B10.BR TCL and by a Th1 KLH-specific, Ek-restricted B10.BR T cell clone.The nature of the downregulatory activity of B10.A APC on IFN-gamma and TNF-alpha production was explored. 2-hour supernatants from antigen-activated B10.A APC/TCL cultures or from B10.A APC activated by LPS had the same inhibitory effects on IFN-gamma and TNF-alpha production by B10.BR TCL.The downregulatory effects of B10.A APC are independent of TNF-alpha, IL-4, IL-10, IL-12p40, IFN-gamma, IL-13, TGF-beta, and PGE2.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Stanford University, California 94305-5020, USA.

ABSTRACT
Development of T helper cell (Th)1 or Th2 cytokine responses is essential for effector and regulatory functions of T helper cells. We have compared cytokine profiles of myelin basic protein (MBP) Ac1-16 peptide-specific T helper cells from inbred mouse strains expressing identical k haplotype-derived MHC class II molecules B10.A and B10.BR, B10.BR T cell lines (TCL) produced Th1 cytokines (including high levels of TNF-alpha) and induced experimental autoimmune encephalomyelitis after adoptive transfer. In contrast, B10.A TCL produced Th2 cytokines (including low levels of TNF-alpha) and were poorly encephalitogenic. The contributions of the genetic origin of the T cells and the APC were explored. Serial restimulations of the B10.BR TCL with B10.A or (B10.A x B10.BR) F1 splenic antigen presenting cells (APC) during the establishment of TCL markedly reduced both Th1 cytokine production and encephalitogenicity. In addition, a single restimulation with B10. A splenic APC reduced IFN-gamma and TNF-alpha production by established Th1 MBP-specific Ak-restricted B10.BR TCL and by a Th1 KLH-specific, Ek-restricted B10.BR T cell clone. These studies suggest that B10.A and B10.BR APC differ in their ability to stimulate IFN-gamma and TNF-alpha production by mature Th1 cells and also influence their Th1/Th2 commitment in vivo. The nature of the downregulatory activity of B10.A APC on IFN-gamma and TNF-alpha production was explored. 2-hour supernatants from antigen-activated B10.A APC/TCL cultures or from B10.A APC activated by LPS had the same inhibitory effects on IFN-gamma and TNF-alpha production by B10.BR TCL. The downregulatory effects of B10.A APC are independent of TNF-alpha, IL-4, IL-10, IL-12p40, IFN-gamma, IL-13, TGF-beta, and PGE2. Thus, genetic difference(s) between B10.A and B10.BR APC appear(s) to control the production or activity of a novel soluble cytokine regulatory factor that influences Th1/Th2 commitment and controls production of IFN-gamma and TNF-alpha by mature Th1 cells.

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Downregulation of B10.BR TCL IFN-γ and TNF-α production by B10.A APC or by 2-h supernatant from activated B10.A  APC. (A–C) Independent experiments with MBP-specific B10.BR TCL  3 (A and B) and TCL 4. TCL were restimulated for 24 h in the presence  of B10.BR or B10.A splenic APC, or splenic APC depleted of T cells (A,  lower bars). In some cultures, B10.BR TCL were restimulated with  B10.BR APC and MBP peptide in the presence of 2-h supernatant fluids  from cultures of B10.BR or B10.A TCL stimulated with syngeneic APC  and MBP peptide (B, bottom bars; C, middle bars), or in the presence of 2-h  supernatants from B10.BR or B10.A spleen cells stimulated with LPS (C,  bottom bars). Data presented are means and standard deviations of triplicate  wells for each sample (n = 3). All experiments were repeated at least three  times with similar results.
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Figure 3: Downregulation of B10.BR TCL IFN-γ and TNF-α production by B10.A APC or by 2-h supernatant from activated B10.A APC. (A–C) Independent experiments with MBP-specific B10.BR TCL 3 (A and B) and TCL 4. TCL were restimulated for 24 h in the presence of B10.BR or B10.A splenic APC, or splenic APC depleted of T cells (A, lower bars). In some cultures, B10.BR TCL were restimulated with B10.BR APC and MBP peptide in the presence of 2-h supernatant fluids from cultures of B10.BR or B10.A TCL stimulated with syngeneic APC and MBP peptide (B, bottom bars; C, middle bars), or in the presence of 2-h supernatants from B10.BR or B10.A spleen cells stimulated with LPS (C, bottom bars). Data presented are means and standard deviations of triplicate wells for each sample (n = 3). All experiments were repeated at least three times with similar results.

Mentions: To determine whether a single restimulation with B10.A APC can alter the production of IFN-γ and TNF-α by established B10.BR-derived Th1s, two independently derived Th1 MBP-specific B10.BR TCL (TCL 3 and TCL 4, both of which had been maintained with syngeneic B10.BR APC) were restimulated overnight with B10.BR or B10.A APC before harvesting supernatants for cytokine assays. As shown in Fig. 3, A–C (top bars), IFN-γ and TNF-α levels were downregulated by 40–60% when B10.A, instead of B10.BR, APC were used for the single restimulation. Similar downregulation of IFN-γ and TNF-α was observed when B10.A (but not B10.BR) T cell–depleted splenic APC were used to restimulate B10.BR TCL (Fig. 3 A, bottom bars). Irrespective of the genetic source of APC, IL-2 levels and proliferative responses remained high and IL-4 was not induced (not shown).


Novel genetic regulation of T helper 1 (Th1)/Th2 cytokine production and encephalitogenicity in inbred mouse strains.

Conboy IM, DeKruyff RH, Tate KM, Cao ZA, Moore TA, Umetsu DT, Jones PP - J. Exp. Med. (1997)

Downregulation of B10.BR TCL IFN-γ and TNF-α production by B10.A APC or by 2-h supernatant from activated B10.A  APC. (A–C) Independent experiments with MBP-specific B10.BR TCL  3 (A and B) and TCL 4. TCL were restimulated for 24 h in the presence  of B10.BR or B10.A splenic APC, or splenic APC depleted of T cells (A,  lower bars). In some cultures, B10.BR TCL were restimulated with  B10.BR APC and MBP peptide in the presence of 2-h supernatant fluids  from cultures of B10.BR or B10.A TCL stimulated with syngeneic APC  and MBP peptide (B, bottom bars; C, middle bars), or in the presence of 2-h  supernatants from B10.BR or B10.A spleen cells stimulated with LPS (C,  bottom bars). Data presented are means and standard deviations of triplicate  wells for each sample (n = 3). All experiments were repeated at least three  times with similar results.
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Related In: Results  -  Collection

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Figure 3: Downregulation of B10.BR TCL IFN-γ and TNF-α production by B10.A APC or by 2-h supernatant from activated B10.A APC. (A–C) Independent experiments with MBP-specific B10.BR TCL 3 (A and B) and TCL 4. TCL were restimulated for 24 h in the presence of B10.BR or B10.A splenic APC, or splenic APC depleted of T cells (A, lower bars). In some cultures, B10.BR TCL were restimulated with B10.BR APC and MBP peptide in the presence of 2-h supernatant fluids from cultures of B10.BR or B10.A TCL stimulated with syngeneic APC and MBP peptide (B, bottom bars; C, middle bars), or in the presence of 2-h supernatants from B10.BR or B10.A spleen cells stimulated with LPS (C, bottom bars). Data presented are means and standard deviations of triplicate wells for each sample (n = 3). All experiments were repeated at least three times with similar results.
Mentions: To determine whether a single restimulation with B10.A APC can alter the production of IFN-γ and TNF-α by established B10.BR-derived Th1s, two independently derived Th1 MBP-specific B10.BR TCL (TCL 3 and TCL 4, both of which had been maintained with syngeneic B10.BR APC) were restimulated overnight with B10.BR or B10.A APC before harvesting supernatants for cytokine assays. As shown in Fig. 3, A–C (top bars), IFN-γ and TNF-α levels were downregulated by 40–60% when B10.A, instead of B10.BR, APC were used for the single restimulation. Similar downregulation of IFN-γ and TNF-α was observed when B10.A (but not B10.BR) T cell–depleted splenic APC were used to restimulate B10.BR TCL (Fig. 3 A, bottom bars). Irrespective of the genetic source of APC, IL-2 levels and proliferative responses remained high and IL-4 was not induced (not shown).

Bottom Line: A splenic APC reduced IFN-gamma and TNF-alpha production by established Th1 MBP-specific Ak-restricted B10.BR TCL and by a Th1 KLH-specific, Ek-restricted B10.BR T cell clone.The nature of the downregulatory activity of B10.A APC on IFN-gamma and TNF-alpha production was explored. 2-hour supernatants from antigen-activated B10.A APC/TCL cultures or from B10.A APC activated by LPS had the same inhibitory effects on IFN-gamma and TNF-alpha production by B10.BR TCL.The downregulatory effects of B10.A APC are independent of TNF-alpha, IL-4, IL-10, IL-12p40, IFN-gamma, IL-13, TGF-beta, and PGE2.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Stanford University, California 94305-5020, USA.

ABSTRACT
Development of T helper cell (Th)1 or Th2 cytokine responses is essential for effector and regulatory functions of T helper cells. We have compared cytokine profiles of myelin basic protein (MBP) Ac1-16 peptide-specific T helper cells from inbred mouse strains expressing identical k haplotype-derived MHC class II molecules B10.A and B10.BR, B10.BR T cell lines (TCL) produced Th1 cytokines (including high levels of TNF-alpha) and induced experimental autoimmune encephalomyelitis after adoptive transfer. In contrast, B10.A TCL produced Th2 cytokines (including low levels of TNF-alpha) and were poorly encephalitogenic. The contributions of the genetic origin of the T cells and the APC were explored. Serial restimulations of the B10.BR TCL with B10.A or (B10.A x B10.BR) F1 splenic antigen presenting cells (APC) during the establishment of TCL markedly reduced both Th1 cytokine production and encephalitogenicity. In addition, a single restimulation with B10. A splenic APC reduced IFN-gamma and TNF-alpha production by established Th1 MBP-specific Ak-restricted B10.BR TCL and by a Th1 KLH-specific, Ek-restricted B10.BR T cell clone. These studies suggest that B10.A and B10.BR APC differ in their ability to stimulate IFN-gamma and TNF-alpha production by mature Th1 cells and also influence their Th1/Th2 commitment in vivo. The nature of the downregulatory activity of B10.A APC on IFN-gamma and TNF-alpha production was explored. 2-hour supernatants from antigen-activated B10.A APC/TCL cultures or from B10.A APC activated by LPS had the same inhibitory effects on IFN-gamma and TNF-alpha production by B10.BR TCL. The downregulatory effects of B10.A APC are independent of TNF-alpha, IL-4, IL-10, IL-12p40, IFN-gamma, IL-13, TGF-beta, and PGE2. Thus, genetic difference(s) between B10.A and B10.BR APC appear(s) to control the production or activity of a novel soluble cytokine regulatory factor that influences Th1/Th2 commitment and controls production of IFN-gamma and TNF-alpha by mature Th1 cells.

Show MeSH
Related in: MedlinePlus