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A role for CD4 in peripheral T cell differentiation.

Brown DR, Moskowitz NH, Killeen N, Reiner SL - J. Exp. Med. (1997)

Bottom Line: CD4 serves a prominent role in potentiating antigen recognition by helper T cells.Together, these results demonstrate a previously undescribed role of the CD4 molecule.The requirement for CD4 in Th2 maturation reflects the importance of molecules other than cytokines in the control of helper T cell differentiation.

View Article: PubMed Central - PubMed

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

ABSTRACT
Naive CD4+ T helper cells (Th) differentiate into one of two well-defined cell types during immune responses. Mature Th1 and Th2 cells regulate the type of response as a consequence of the unique cytokines that they secrete. CD4 serves a prominent role in potentiating antigen recognition by helper T cells. We have examined the role of CD4 in peripheral T cell differentiation by studying helper T cells from mice with a congenital defect in CD4 expression. After protein immunization or infection with Leishmania major, CD4-deficient mice were incapable of mounting antigen-specific Th2 responses, but retained their Th1 potency. CD4-deficient, T cell receptor transgenic T cells were also incapable of Th2 differentiation after in vitro activation. Expression of a wild-type CD4 transgene corrected the Th2 defect of CD4-deficient mice in all immune responses tested. To investigate the role of the cytoplasmic domain, mice reconstituted with a truncated CD4 molecule were also studied. Expression of the tailless CD4 transgene could not rescue the Th2 defect of CD4-deficient mice immunized with protein or CD4-deficient transgenic T cells activated in vitro, raising the possibility that the cytoplasmic domain of CD4 may influence Th2 generation. Expression of the tailless transgene was, however, capable of restoring Th2 development in CD4-deficient mice infected with L. major or CD4-deficient transgenic T cells activated in the presence of recombinant IL-4, demonstrating that the cytoplasmic domain is not absolutely required for Th2 development. Together, these results demonstrate a previously undescribed role of the CD4 molecule. The requirement for CD4 in Th2 maturation reflects the importance of molecules other than cytokines in the control of helper T cell differentiation.

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The absence of CD4 impairs Th2 differentiation when T  cells are primed without APCs. (A) Purified CD8− T cells were isolated  from lymph nodes of CD4+ or CD4° mice, and 5 × 105 cells were plated  in wells precoated with anti-CD3 (10 μg/ml) plus rIL-4. After 5 d, cells  were washed extensively and 105 cells were restimulated on anti-CD3– coated plates in triplicate for an additional 48 h. IL-4 (filled bars) and IFN-γ  (open bars) were quantitated from supernatants by ELISA. The results represent the mean of triplicate determinations and are representative of six  separate experiments. (B) Purified helper lineage T cells were primed as  described in A, using varying numbers of cells in the primary stimulation  (x-axis) and 2 × 105 cells/ml in the secondary stimulation (B, top), or 2.5 ×  106 cells/ml in the primary stimulation and varying numbers of cells in  the secondary stimulation (x-axis; B, bottom). rIL-4 was withheld (B, left)  or added (B, right) to the primary cultures where indicated. (C) CD8− T  cells isolated from lymph nodes of CD4+ or CD4° mice were cultured on  plates precoated with the indicated concentration of anti-CD3. After 48 h,  [3H]thymidine was added, and plates were harvested 12–14 h later. The  mean of triplicate wells was used to determine the percent maximum thymidine incorporation for each group.
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Figure 4: The absence of CD4 impairs Th2 differentiation when T cells are primed without APCs. (A) Purified CD8− T cells were isolated from lymph nodes of CD4+ or CD4° mice, and 5 × 105 cells were plated in wells precoated with anti-CD3 (10 μg/ml) plus rIL-4. After 5 d, cells were washed extensively and 105 cells were restimulated on anti-CD3– coated plates in triplicate for an additional 48 h. IL-4 (filled bars) and IFN-γ (open bars) were quantitated from supernatants by ELISA. The results represent the mean of triplicate determinations and are representative of six separate experiments. (B) Purified helper lineage T cells were primed as described in A, using varying numbers of cells in the primary stimulation (x-axis) and 2 × 105 cells/ml in the secondary stimulation (B, top), or 2.5 × 106 cells/ml in the primary stimulation and varying numbers of cells in the secondary stimulation (x-axis; B, bottom). rIL-4 was withheld (B, left) or added (B, right) to the primary cultures where indicated. (C) CD8− T cells isolated from lymph nodes of CD4+ or CD4° mice were cultured on plates precoated with the indicated concentration of anti-CD3. After 48 h, [3H]thymidine was added, and plates were harvested 12–14 h later. The mean of triplicate wells was used to determine the percent maximum thymidine incorporation for each group.

Mentions: As a further test of the intrinsic differentiation capacity of CD4-deficient helper T cells, we established an antigen– MHC-independent system of T cell activation thereby eliminating differences in T cell–APC adhesion of CD4+ and CD4° T cells. Highly purified CD8− helper T cells were stimulated with plate-bound anti-CD3 mAb plus rIL-4 for 5 d. Cells were then washed and restimulated with plate-bound anti-CD3 to assess their maturational state. CD4+ T cells were capable of significant IL-4 and modest IFN-γ production, whereas the T cells from CD4° mice produced high amounts of IFN-γ with negligible IL-4 (Fig. 4 A). We also attempted to augment the efficiency of Th2 maturation or IL-4 secretion by increasing the density of T cells in the primary or secondary cultures, respectively. For CD4+ T cells, increasing cell number in the primary culture was sufficient to promote Th2 maturation, even in the absence of rIL-4 (Fig. 4 B). For CD4° T cells, even the highest cell number plus the addition of rIL-4 failed to elicit IL-4 production (Fig. 4 B). Moreover, neither the addition of rIL-2 nor co-culture of CD4+ with CD4° T cells could rescue Th2 differentiation in the CD4° population (data not shown).


A role for CD4 in peripheral T cell differentiation.

Brown DR, Moskowitz NH, Killeen N, Reiner SL - J. Exp. Med. (1997)

The absence of CD4 impairs Th2 differentiation when T  cells are primed without APCs. (A) Purified CD8− T cells were isolated  from lymph nodes of CD4+ or CD4° mice, and 5 × 105 cells were plated  in wells precoated with anti-CD3 (10 μg/ml) plus rIL-4. After 5 d, cells  were washed extensively and 105 cells were restimulated on anti-CD3– coated plates in triplicate for an additional 48 h. IL-4 (filled bars) and IFN-γ  (open bars) were quantitated from supernatants by ELISA. The results represent the mean of triplicate determinations and are representative of six  separate experiments. (B) Purified helper lineage T cells were primed as  described in A, using varying numbers of cells in the primary stimulation  (x-axis) and 2 × 105 cells/ml in the secondary stimulation (B, top), or 2.5 ×  106 cells/ml in the primary stimulation and varying numbers of cells in  the secondary stimulation (x-axis; B, bottom). rIL-4 was withheld (B, left)  or added (B, right) to the primary cultures where indicated. (C) CD8− T  cells isolated from lymph nodes of CD4+ or CD4° mice were cultured on  plates precoated with the indicated concentration of anti-CD3. After 48 h,  [3H]thymidine was added, and plates were harvested 12–14 h later. The  mean of triplicate wells was used to determine the percent maximum thymidine incorporation for each group.
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Figure 4: The absence of CD4 impairs Th2 differentiation when T cells are primed without APCs. (A) Purified CD8− T cells were isolated from lymph nodes of CD4+ or CD4° mice, and 5 × 105 cells were plated in wells precoated with anti-CD3 (10 μg/ml) plus rIL-4. After 5 d, cells were washed extensively and 105 cells were restimulated on anti-CD3– coated plates in triplicate for an additional 48 h. IL-4 (filled bars) and IFN-γ (open bars) were quantitated from supernatants by ELISA. The results represent the mean of triplicate determinations and are representative of six separate experiments. (B) Purified helper lineage T cells were primed as described in A, using varying numbers of cells in the primary stimulation (x-axis) and 2 × 105 cells/ml in the secondary stimulation (B, top), or 2.5 × 106 cells/ml in the primary stimulation and varying numbers of cells in the secondary stimulation (x-axis; B, bottom). rIL-4 was withheld (B, left) or added (B, right) to the primary cultures where indicated. (C) CD8− T cells isolated from lymph nodes of CD4+ or CD4° mice were cultured on plates precoated with the indicated concentration of anti-CD3. After 48 h, [3H]thymidine was added, and plates were harvested 12–14 h later. The mean of triplicate wells was used to determine the percent maximum thymidine incorporation for each group.
Mentions: As a further test of the intrinsic differentiation capacity of CD4-deficient helper T cells, we established an antigen– MHC-independent system of T cell activation thereby eliminating differences in T cell–APC adhesion of CD4+ and CD4° T cells. Highly purified CD8− helper T cells were stimulated with plate-bound anti-CD3 mAb plus rIL-4 for 5 d. Cells were then washed and restimulated with plate-bound anti-CD3 to assess their maturational state. CD4+ T cells were capable of significant IL-4 and modest IFN-γ production, whereas the T cells from CD4° mice produced high amounts of IFN-γ with negligible IL-4 (Fig. 4 A). We also attempted to augment the efficiency of Th2 maturation or IL-4 secretion by increasing the density of T cells in the primary or secondary cultures, respectively. For CD4+ T cells, increasing cell number in the primary culture was sufficient to promote Th2 maturation, even in the absence of rIL-4 (Fig. 4 B). For CD4° T cells, even the highest cell number plus the addition of rIL-4 failed to elicit IL-4 production (Fig. 4 B). Moreover, neither the addition of rIL-2 nor co-culture of CD4+ with CD4° T cells could rescue Th2 differentiation in the CD4° population (data not shown).

Bottom Line: CD4 serves a prominent role in potentiating antigen recognition by helper T cells.Together, these results demonstrate a previously undescribed role of the CD4 molecule.The requirement for CD4 in Th2 maturation reflects the importance of molecules other than cytokines in the control of helper T cell differentiation.

View Article: PubMed Central - PubMed

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

ABSTRACT
Naive CD4+ T helper cells (Th) differentiate into one of two well-defined cell types during immune responses. Mature Th1 and Th2 cells regulate the type of response as a consequence of the unique cytokines that they secrete. CD4 serves a prominent role in potentiating antigen recognition by helper T cells. We have examined the role of CD4 in peripheral T cell differentiation by studying helper T cells from mice with a congenital defect in CD4 expression. After protein immunization or infection with Leishmania major, CD4-deficient mice were incapable of mounting antigen-specific Th2 responses, but retained their Th1 potency. CD4-deficient, T cell receptor transgenic T cells were also incapable of Th2 differentiation after in vitro activation. Expression of a wild-type CD4 transgene corrected the Th2 defect of CD4-deficient mice in all immune responses tested. To investigate the role of the cytoplasmic domain, mice reconstituted with a truncated CD4 molecule were also studied. Expression of the tailless CD4 transgene could not rescue the Th2 defect of CD4-deficient mice immunized with protein or CD4-deficient transgenic T cells activated in vitro, raising the possibility that the cytoplasmic domain of CD4 may influence Th2 generation. Expression of the tailless transgene was, however, capable of restoring Th2 development in CD4-deficient mice infected with L. major or CD4-deficient transgenic T cells activated in the presence of recombinant IL-4, demonstrating that the cytoplasmic domain is not absolutely required for Th2 development. Together, these results demonstrate a previously undescribed role of the CD4 molecule. The requirement for CD4 in Th2 maturation reflects the importance of molecules other than cytokines in the control of helper T cell differentiation.

Show MeSH
Related in: MedlinePlus