Limits...
T helper subset differentiation in the absence of invariant chain.

Brown DR, Swier K, Moskowitz NH, Naujokas MF, Locksley RM, Reiner SL - J. Exp. Med. (1997)

Bottom Line: We wished to determine whether this unexpected efficiency of T helper subset induction despite inefficient T cell stimulation could be modeled in vitro.In the presence of rIL-12 or rIL-4 naive parasite-specific transgenic T cells could mature into IFN-gamma-or IL-4-secreting T helper cells, respectively, even when antigen presentation was suboptimal or antigen dose was submitogenic.These experiments demonstrate that activation of T helper cells to a threshold required for IL-2 production or proliferation is not required to achieve induction of disease-regulating T helper cell effector functions, and that pathogen-associated secondary activation signals may facilitate the full differentiation of T helper subsets during limiting presentation of antigenic peptides.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Gwen Knapp Center for Lupus & Immunology Research, University of Chicago, Illinois 60637, USA.

ABSTRACT
The outcome of murine infection with Leishmania major is regulated by major histocompatibility complex class II-restricted T helper cells. Invariant chain-deficient (Ii -/-) mice have impaired ability to present major histocompatibility complex class II-restricted antigens, and reduced numbers of CD4+ T cells. Despite these deficits, C57BL/6 Ii -/- mice controlled L. major infection comparably to wild-type mice. As assessed by mRNA analysis and in vitro antigen restimulation for IFN-gamma, Ii -/- mice had normal induction of Th1 subset differentiation even though antigen-dependent proliferation of their lymph node cells was substantially compromised. In addition, BALB/c Ii -/- mice exhibited a progressive course of infection and Th2 effector cell development that were comparable to that seen in wild-type BALB/c mice. We wished to determine whether this unexpected efficiency of T helper subset induction despite inefficient T cell stimulation could be modeled in vitro. In the presence of rIL-12 or rIL-4 naive parasite-specific transgenic T cells could mature into IFN-gamma-or IL-4-secreting T helper cells, respectively, even when antigen presentation was suboptimal or antigen dose was submitogenic. These experiments demonstrate that activation of T helper cells to a threshold required for IL-2 production or proliferation is not required to achieve induction of disease-regulating T helper cell effector functions, and that pathogen-associated secondary activation signals may facilitate the full differentiation of T helper subsets during limiting presentation of antigenic peptides.

Show MeSH

Related in: MedlinePlus

IL-4 supports Th2 differentiation by T cells primed on Ii −/−  APCs. CD4-selected TCR transgenic lymph node cells (2 × 105 cells/ well) were stimulated in a 1 ml volume in 24-well plates with either Ii + or  Ii −/− irradiated H-2d spleen cells (1 × 106 cells/well), plus recombinant  LACK antigen (0.5 μg/ml), with (+) or without (−) rIL-4 (5,000 U/ml)  as designated on the x-axis. 6 d later, viable cells were purified and  washed, and 2 × 105 T cells were restimulated with the same Ii-genotype  APCs (1.2 × 106/well) used in the primary stimulation in the presence of  antigen (0.5 μg/ml), but without rIL-4. After 48 h, supernatants were  collected and assayed for IL-4 by ELISA. Values represent means and  standard deviations of triplicate determinations.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2196096&req=5

Figure 10: IL-4 supports Th2 differentiation by T cells primed on Ii −/− APCs. CD4-selected TCR transgenic lymph node cells (2 × 105 cells/ well) were stimulated in a 1 ml volume in 24-well plates with either Ii + or Ii −/− irradiated H-2d spleen cells (1 × 106 cells/well), plus recombinant LACK antigen (0.5 μg/ml), with (+) or without (−) rIL-4 (5,000 U/ml) as designated on the x-axis. 6 d later, viable cells were purified and washed, and 2 × 105 T cells were restimulated with the same Ii-genotype APCs (1.2 × 106/well) used in the primary stimulation in the presence of antigen (0.5 μg/ml), but without rIL-4. After 48 h, supernatants were collected and assayed for IL-4 by ELISA. Values represent means and standard deviations of triplicate determinations.

Mentions: To confirm that the threshold for Th2 development could be maintained in the absence of invariant chain, LACK-specific transgenic T cells were primed using antigen-pulsed, irradiated spleen cells from Ii + or Ii −/− mice in the presence or absence of rIL-4. After 6 d, viable cells were washed extensively, counted, and redistributed with freshly prepared, antigen-pulsed, irradiated spleen cells from the same Ii backgrounds, but in the absence of rIL-4. After 48 h, the supernatants were analyzed for IL-4 (Fig. 10). Recovery of IL-4 was substantially enhanced during the secondary stimulation when cells were primed using Ii −/− APCs, but only when rIL-4 was included during the primary incubation with LACK.


T helper subset differentiation in the absence of invariant chain.

Brown DR, Swier K, Moskowitz NH, Naujokas MF, Locksley RM, Reiner SL - J. Exp. Med. (1997)

IL-4 supports Th2 differentiation by T cells primed on Ii −/−  APCs. CD4-selected TCR transgenic lymph node cells (2 × 105 cells/ well) were stimulated in a 1 ml volume in 24-well plates with either Ii + or  Ii −/− irradiated H-2d spleen cells (1 × 106 cells/well), plus recombinant  LACK antigen (0.5 μg/ml), with (+) or without (−) rIL-4 (5,000 U/ml)  as designated on the x-axis. 6 d later, viable cells were purified and  washed, and 2 × 105 T cells were restimulated with the same Ii-genotype  APCs (1.2 × 106/well) used in the primary stimulation in the presence of  antigen (0.5 μg/ml), but without rIL-4. After 48 h, supernatants were  collected and assayed for IL-4 by ELISA. Values represent means and  standard deviations of triplicate determinations.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2196096&req=5

Figure 10: IL-4 supports Th2 differentiation by T cells primed on Ii −/− APCs. CD4-selected TCR transgenic lymph node cells (2 × 105 cells/ well) were stimulated in a 1 ml volume in 24-well plates with either Ii + or Ii −/− irradiated H-2d spleen cells (1 × 106 cells/well), plus recombinant LACK antigen (0.5 μg/ml), with (+) or without (−) rIL-4 (5,000 U/ml) as designated on the x-axis. 6 d later, viable cells were purified and washed, and 2 × 105 T cells were restimulated with the same Ii-genotype APCs (1.2 × 106/well) used in the primary stimulation in the presence of antigen (0.5 μg/ml), but without rIL-4. After 48 h, supernatants were collected and assayed for IL-4 by ELISA. Values represent means and standard deviations of triplicate determinations.
Mentions: To confirm that the threshold for Th2 development could be maintained in the absence of invariant chain, LACK-specific transgenic T cells were primed using antigen-pulsed, irradiated spleen cells from Ii + or Ii −/− mice in the presence or absence of rIL-4. After 6 d, viable cells were washed extensively, counted, and redistributed with freshly prepared, antigen-pulsed, irradiated spleen cells from the same Ii backgrounds, but in the absence of rIL-4. After 48 h, the supernatants were analyzed for IL-4 (Fig. 10). Recovery of IL-4 was substantially enhanced during the secondary stimulation when cells were primed using Ii −/− APCs, but only when rIL-4 was included during the primary incubation with LACK.

Bottom Line: We wished to determine whether this unexpected efficiency of T helper subset induction despite inefficient T cell stimulation could be modeled in vitro.In the presence of rIL-12 or rIL-4 naive parasite-specific transgenic T cells could mature into IFN-gamma-or IL-4-secreting T helper cells, respectively, even when antigen presentation was suboptimal or antigen dose was submitogenic.These experiments demonstrate that activation of T helper cells to a threshold required for IL-2 production or proliferation is not required to achieve induction of disease-regulating T helper cell effector functions, and that pathogen-associated secondary activation signals may facilitate the full differentiation of T helper subsets during limiting presentation of antigenic peptides.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Gwen Knapp Center for Lupus & Immunology Research, University of Chicago, Illinois 60637, USA.

ABSTRACT
The outcome of murine infection with Leishmania major is regulated by major histocompatibility complex class II-restricted T helper cells. Invariant chain-deficient (Ii -/-) mice have impaired ability to present major histocompatibility complex class II-restricted antigens, and reduced numbers of CD4+ T cells. Despite these deficits, C57BL/6 Ii -/- mice controlled L. major infection comparably to wild-type mice. As assessed by mRNA analysis and in vitro antigen restimulation for IFN-gamma, Ii -/- mice had normal induction of Th1 subset differentiation even though antigen-dependent proliferation of their lymph node cells was substantially compromised. In addition, BALB/c Ii -/- mice exhibited a progressive course of infection and Th2 effector cell development that were comparable to that seen in wild-type BALB/c mice. We wished to determine whether this unexpected efficiency of T helper subset induction despite inefficient T cell stimulation could be modeled in vitro. In the presence of rIL-12 or rIL-4 naive parasite-specific transgenic T cells could mature into IFN-gamma-or IL-4-secreting T helper cells, respectively, even when antigen presentation was suboptimal or antigen dose was submitogenic. These experiments demonstrate that activation of T helper cells to a threshold required for IL-2 production or proliferation is not required to achieve induction of disease-regulating T helper cell effector functions, and that pathogen-associated secondary activation signals may facilitate the full differentiation of T helper subsets during limiting presentation of antigenic peptides.

Show MeSH
Related in: MedlinePlus