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Interleukin 4-producing CD4 T cells arise from different precursors depending on the conditions of antigen exposure in vivo.

Foucras G, Gapin L, Coureau C, Kanellopoulos JM, Guéry JC - J. Exp. Med. (2000)

Bottom Line: We show that public T cell clones are predominantly associated with type 1 or 2 effector Th cells recovered after primary immunization in complete or incomplete Freund's adjuvant, respectively.Conversely, continuous administration of soluble antigen, which induces strong memory Th2 response, is associated with a dose-dependent reduction of public clone size by a mechanism resembling clonal anergy.Thus, soluble HEL-induced Th2 cells do not express the public complementarity determining region 3 motifs characteristic of immunogenic challenge in the presence of adjuvant.

View Article: PubMed Central - PubMed

Affiliation: Institut National de la Santé et de la Recherche Médicale (INSERM) U28, Institut Fédératif de Recherche 30, Hôpital Purpan, 31059 Toulouse Cedex, France.

ABSTRACT
The precursor origin of T helper (Th) cell subsets in vivo has been difficult to study and remains poorly investigated. We have previously shown that chronic administration of soluble protein antigen induces selective development of antigen-specific CD4 Th2 cells in genetically predisposed mouse strains. To analyze the origin of effector T cells in this model, we designed a competitive polymerase chain reaction-based approach to track public BV-J rearrangement expressed by CD4 T cells specific for hen egg white lysozyme (HEL) in BALB/c mice. We show that public T cell clones are predominantly associated with type 1 or 2 effector Th cells recovered after primary immunization in complete or incomplete Freund's adjuvant, respectively. Conversely, continuous administration of soluble antigen, which induces strong memory Th2 response, is associated with a dose-dependent reduction of public clone size by a mechanism resembling clonal anergy. Thus, soluble HEL-induced Th2 cells do not express the public complementarity determining region 3 motifs characteristic of immunogenic challenge in the presence of adjuvant. These results demonstrate that there are multiple pathways of induction of Th2 responses depending on the condition of antigen exposure in vivo, i.e., clonal immune deviation versus recruitment of a different pool of precursor cells.

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Immunoscope analysis reveals a dose-dependent loss of public repertoire BV8S2-J1S5HEL 107–116 in soluble HEL–induced Th2 cells. Popliteal LNCs from mice untreated or treated with the indicated amounts of HEL in pump were stimulated in vitro with HEL (10 μM). (A) After 4 d, CD4 T cells were purified and subjected to RNA extraction. PCR amplification of BV8S2-BC rearrangements on cDNA and Immunoscope analysis were then performed as described in Materials and Methods. Data are from one representative mouse out of four analyzed per group. (B) Data from the Immunoscope analysis were expressed as ratio of the area under the public CDR3–specific peak by the sum of the CDR3 peaks obtained in the BC-specific run-off reactions. (C) Ag-specific IFN-γ and IL-4 production was measured in 72-h culture supernatants by ELISA. Data are expressed as mean ± SD of four mice per group. Results are representative of two experiments performed.
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Figure 3: Immunoscope analysis reveals a dose-dependent loss of public repertoire BV8S2-J1S5HEL 107–116 in soluble HEL–induced Th2 cells. Popliteal LNCs from mice untreated or treated with the indicated amounts of HEL in pump were stimulated in vitro with HEL (10 μM). (A) After 4 d, CD4 T cells were purified and subjected to RNA extraction. PCR amplification of BV8S2-BC rearrangements on cDNA and Immunoscope analysis were then performed as described in Materials and Methods. Data are from one representative mouse out of four analyzed per group. (B) Data from the Immunoscope analysis were expressed as ratio of the area under the public CDR3–specific peak by the sum of the CDR3 peaks obtained in the BC-specific run-off reactions. (C) Ag-specific IFN-γ and IL-4 production was measured in 72-h culture supernatants by ELISA. Data are expressed as mean ± SD of four mice per group. Results are representative of two experiments performed.

Mentions: We next tested the effect of pretreatment with a wide dose range of soluble HEL on the expression of the public rearrangement in HEL-specific CD4 T cells. Using the PCR-based approach Immunoscope, we analyzed the CDR3 size distribution of TCRs bearing the BV8S2-J1S5 rearrangement as previously described 16. As shown in Fig. 3 A, the CDR3 size distribution among all BV8S2-bearing cells gives a set of seven peaks, each separated by three nucleotides in control and soluble HEL–injected mice. Conversely, a major expansion with a CDR3 size of eight amino acids is observed with BJ1S5 and public CDR3–specific primers in control animals immunized with HEL–CFA (Fig. 3 A). Interestingly, continuous administration of soluble HEL is associated with an increased heterogeneity in the CDR3 size in this BV-J combination due to a diminished expression of the BV8S2-J1S5HEL 107–116 public rearrangement. Data from four individual mice per group are summarized in Fig. 3 B, where CDR3 BV8S2-J1S5HEL 107–116–specific products are expressed as percent of the sum of the BV8S2-BC peaks. Therefore, continuous administration of low dose soluble HEL results in a strong reduction of Ag-specific T cells expressing the public CDR3 BV8S2-J1S5HEL 107–116 (Fig. 3 B), which can be correlated with the downregulation of Th1 response but not with the expansion of IL-4–producing cells (Fig. 3 C).


Interleukin 4-producing CD4 T cells arise from different precursors depending on the conditions of antigen exposure in vivo.

Foucras G, Gapin L, Coureau C, Kanellopoulos JM, Guéry JC - J. Exp. Med. (2000)

Immunoscope analysis reveals a dose-dependent loss of public repertoire BV8S2-J1S5HEL 107–116 in soluble HEL–induced Th2 cells. Popliteal LNCs from mice untreated or treated with the indicated amounts of HEL in pump were stimulated in vitro with HEL (10 μM). (A) After 4 d, CD4 T cells were purified and subjected to RNA extraction. PCR amplification of BV8S2-BC rearrangements on cDNA and Immunoscope analysis were then performed as described in Materials and Methods. Data are from one representative mouse out of four analyzed per group. (B) Data from the Immunoscope analysis were expressed as ratio of the area under the public CDR3–specific peak by the sum of the CDR3 peaks obtained in the BC-specific run-off reactions. (C) Ag-specific IFN-γ and IL-4 production was measured in 72-h culture supernatants by ELISA. Data are expressed as mean ± SD of four mice per group. Results are representative of two experiments performed.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2195842&req=5

Figure 3: Immunoscope analysis reveals a dose-dependent loss of public repertoire BV8S2-J1S5HEL 107–116 in soluble HEL–induced Th2 cells. Popliteal LNCs from mice untreated or treated with the indicated amounts of HEL in pump were stimulated in vitro with HEL (10 μM). (A) After 4 d, CD4 T cells were purified and subjected to RNA extraction. PCR amplification of BV8S2-BC rearrangements on cDNA and Immunoscope analysis were then performed as described in Materials and Methods. Data are from one representative mouse out of four analyzed per group. (B) Data from the Immunoscope analysis were expressed as ratio of the area under the public CDR3–specific peak by the sum of the CDR3 peaks obtained in the BC-specific run-off reactions. (C) Ag-specific IFN-γ and IL-4 production was measured in 72-h culture supernatants by ELISA. Data are expressed as mean ± SD of four mice per group. Results are representative of two experiments performed.
Mentions: We next tested the effect of pretreatment with a wide dose range of soluble HEL on the expression of the public rearrangement in HEL-specific CD4 T cells. Using the PCR-based approach Immunoscope, we analyzed the CDR3 size distribution of TCRs bearing the BV8S2-J1S5 rearrangement as previously described 16. As shown in Fig. 3 A, the CDR3 size distribution among all BV8S2-bearing cells gives a set of seven peaks, each separated by three nucleotides in control and soluble HEL–injected mice. Conversely, a major expansion with a CDR3 size of eight amino acids is observed with BJ1S5 and public CDR3–specific primers in control animals immunized with HEL–CFA (Fig. 3 A). Interestingly, continuous administration of soluble HEL is associated with an increased heterogeneity in the CDR3 size in this BV-J combination due to a diminished expression of the BV8S2-J1S5HEL 107–116 public rearrangement. Data from four individual mice per group are summarized in Fig. 3 B, where CDR3 BV8S2-J1S5HEL 107–116–specific products are expressed as percent of the sum of the BV8S2-BC peaks. Therefore, continuous administration of low dose soluble HEL results in a strong reduction of Ag-specific T cells expressing the public CDR3 BV8S2-J1S5HEL 107–116 (Fig. 3 B), which can be correlated with the downregulation of Th1 response but not with the expansion of IL-4–producing cells (Fig. 3 C).

Bottom Line: We show that public T cell clones are predominantly associated with type 1 or 2 effector Th cells recovered after primary immunization in complete or incomplete Freund's adjuvant, respectively.Conversely, continuous administration of soluble antigen, which induces strong memory Th2 response, is associated with a dose-dependent reduction of public clone size by a mechanism resembling clonal anergy.Thus, soluble HEL-induced Th2 cells do not express the public complementarity determining region 3 motifs characteristic of immunogenic challenge in the presence of adjuvant.

View Article: PubMed Central - PubMed

Affiliation: Institut National de la Santé et de la Recherche Médicale (INSERM) U28, Institut Fédératif de Recherche 30, Hôpital Purpan, 31059 Toulouse Cedex, France.

ABSTRACT
The precursor origin of T helper (Th) cell subsets in vivo has been difficult to study and remains poorly investigated. We have previously shown that chronic administration of soluble protein antigen induces selective development of antigen-specific CD4 Th2 cells in genetically predisposed mouse strains. To analyze the origin of effector T cells in this model, we designed a competitive polymerase chain reaction-based approach to track public BV-J rearrangement expressed by CD4 T cells specific for hen egg white lysozyme (HEL) in BALB/c mice. We show that public T cell clones are predominantly associated with type 1 or 2 effector Th cells recovered after primary immunization in complete or incomplete Freund's adjuvant, respectively. Conversely, continuous administration of soluble antigen, which induces strong memory Th2 response, is associated with a dose-dependent reduction of public clone size by a mechanism resembling clonal anergy. Thus, soluble HEL-induced Th2 cells do not express the public complementarity determining region 3 motifs characteristic of immunogenic challenge in the presence of adjuvant. These results demonstrate that there are multiple pathways of induction of Th2 responses depending on the condition of antigen exposure in vivo, i.e., clonal immune deviation versus recruitment of a different pool of precursor cells.

Show MeSH
Related in: MedlinePlus