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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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The public repertoire BV8S2-J1S5 specific for the dominant I-Ed–restricted HEL 107–116 determinant is associated with IFN-γ–producing cells. (A) After 6 d of in vitro stimulation, purified CD4 T cells from HEL–CFA-immunized mice were stained for intracytoplasmic synthesis of IFN-γ and IL-4 as described in Fig. 1 and isolated by FACS® as indicated. (B) Semiquantitative PCR for public rearrangement BV8S2-J1S5, normalized on constant region BC gene amplification, was performed on DNA from whole CD4, IFN-γ−, and IFN-γ+ cells. (C) Run-off reactions were performed on PCR products with a CDR3-specific fluorescent primer to confirm the presence of BV8S2-J1S5HEL 107–116–specific sequence. Results are representative of three experiments performed.
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Figure 2: The public repertoire BV8S2-J1S5 specific for the dominant I-Ed–restricted HEL 107–116 determinant is associated with IFN-γ–producing cells. (A) After 6 d of in vitro stimulation, purified CD4 T cells from HEL–CFA-immunized mice were stained for intracytoplasmic synthesis of IFN-γ and IL-4 as described in Fig. 1 and isolated by FACS® as indicated. (B) Semiquantitative PCR for public rearrangement BV8S2-J1S5, normalized on constant region BC gene amplification, was performed on DNA from whole CD4, IFN-γ−, and IFN-γ+ cells. (C) Run-off reactions were performed on PCR products with a CDR3-specific fluorescent primer to confirm the presence of BV8S2-J1S5HEL 107–116–specific sequence. Results are representative of three experiments performed.

Mentions: It was previously shown that the T cell response against the immunodominant HEL 107–116 epitope in H-2d mice involves a public BV8S2-J1S5 repertoire found in all animals with a specific CDR3 amino acid sequence, GTGNNQAP 16. Immune LNCs from HEL–CFA-primed mice secrete IFN-γ in response to both HEL protein and HEL 103–117 peptide. To analyze whether these IFN-γ–producing cells express the public rearrangement, we analyzed the presence of the public CDR3 motif (BV8S2-J1S5HEL 107–116) on the DNA of HEL-specific CD4 T cells sorted on the basis of intracellular expression of IFN-γ. As shown in Fig. 2 A, DNA was extracted from highly purified (>95%) IFN-γ− and IFN-γ+ CD4 T cells from LNCs restimulated in vitro as described in Fig. 1 A and subjected to PCR using BV8S2- and BJ1S5-specific primers. As shown in Fig. 2 B, the rearranged BV8S2-J1S5 products are preferentially amplified from the DNA extracted from IFN-γ+ but not IFN-γ− CD4 T cells. Fig. 2 C shows the run-off reaction performed with CDR3-specific primers, demonstrating that the public CDR3 BV8S2-J1S5HEL 107–116 motif is mainly expressed in IFN-γ–producing CD4 T cells.


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)

The public repertoire BV8S2-J1S5 specific for the dominant I-Ed–restricted HEL 107–116 determinant is associated with IFN-γ–producing cells. (A) After 6 d of in vitro stimulation, purified CD4 T cells from HEL–CFA-immunized mice were stained for intracytoplasmic synthesis of IFN-γ and IL-4 as described in Fig. 1 and isolated by FACS® as indicated. (B) Semiquantitative PCR for public rearrangement BV8S2-J1S5, normalized on constant region BC gene amplification, was performed on DNA from whole CD4, IFN-γ−, and IFN-γ+ cells. (C) Run-off reactions were performed on PCR products with a CDR3-specific fluorescent primer to confirm the presence of BV8S2-J1S5HEL 107–116–specific sequence. Results are representative of three experiments performed.
© Copyright Policy
Related In: Results  -  Collection

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

Figure 2: The public repertoire BV8S2-J1S5 specific for the dominant I-Ed–restricted HEL 107–116 determinant is associated with IFN-γ–producing cells. (A) After 6 d of in vitro stimulation, purified CD4 T cells from HEL–CFA-immunized mice were stained for intracytoplasmic synthesis of IFN-γ and IL-4 as described in Fig. 1 and isolated by FACS® as indicated. (B) Semiquantitative PCR for public rearrangement BV8S2-J1S5, normalized on constant region BC gene amplification, was performed on DNA from whole CD4, IFN-γ−, and IFN-γ+ cells. (C) Run-off reactions were performed on PCR products with a CDR3-specific fluorescent primer to confirm the presence of BV8S2-J1S5HEL 107–116–specific sequence. Results are representative of three experiments performed.
Mentions: It was previously shown that the T cell response against the immunodominant HEL 107–116 epitope in H-2d mice involves a public BV8S2-J1S5 repertoire found in all animals with a specific CDR3 amino acid sequence, GTGNNQAP 16. Immune LNCs from HEL–CFA-primed mice secrete IFN-γ in response to both HEL protein and HEL 103–117 peptide. To analyze whether these IFN-γ–producing cells express the public rearrangement, we analyzed the presence of the public CDR3 motif (BV8S2-J1S5HEL 107–116) on the DNA of HEL-specific CD4 T cells sorted on the basis of intracellular expression of IFN-γ. As shown in Fig. 2 A, DNA was extracted from highly purified (>95%) IFN-γ− and IFN-γ+ CD4 T cells from LNCs restimulated in vitro as described in Fig. 1 A and subjected to PCR using BV8S2- and BJ1S5-specific primers. As shown in Fig. 2 B, the rearranged BV8S2-J1S5 products are preferentially amplified from the DNA extracted from IFN-γ+ but not IFN-γ− CD4 T cells. Fig. 2 C shows the run-off reaction performed with CDR3-specific primers, demonstrating that the public CDR3 BV8S2-J1S5HEL 107–116 motif is mainly expressed in IFN-γ–producing CD4 T cells.

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