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Differential expression of chemokine receptors and chemotactic responsiveness of type 1 T helper cells (Th1s) and Th2s.

Bonecchi R, Bianchi G, Bordignon PP, D'Ambrosio D, Lang R, Borsatti A, Sozzani S, Allavena P, Gray PA, Mantovani A, Sinigaglia F - J. Exp. Med. (1998)

Bottom Line: In agreement with the differential chemokine receptor expression, Th1s and Th2s selectively migrate in response to the corresponding chemokines.The differential expression of chemokine receptors may dictate, to a large extent, the migration and tissue homing of Th1s and Th2s.It may also determine different susceptibility of Th1s and Th2s to human immunodeficiency virus strains using different fusion coreceptors.

View Article: PubMed Central - PubMed

Affiliation: Istituto di Ricerche Farmacologiche "Mario Negri", I-20157 Milan, Italy.

ABSTRACT
T helper cells type 1 (Th1s) that produce interferon-gamma predominantly mediate cellular immune responses and are involved in the development of chronic inflammatory conditions, whereas Th2s which produce large amounts of IL-4 and IL-5 upregulate IgE production and are prominent in the pathogenesis of allergic diseases. The precise factors determining whether Th1- or Th2-mediated immune responses preferentially occur at a peripheral site of antigen exposure are largely unknown. Chemokines, a superfamily of polypeptide mediators, are a key component of the leukocyte recruitment process. Here we report that among four CXC (CXCR1-4) and five CC (CCR1-5) chemokine receptors analyzed, CXCR3 and CCR5 are preferentially expressed in human Th1s. In contrast, Th2s preferentially express CCR4 and, to a lesser extent, CCR3. In agreement with the differential chemokine receptor expression, Th1s and Th2s selectively migrate in response to the corresponding chemokines. The differential expression of chemokine receptors may dictate, to a large extent, the migration and tissue homing of Th1s and Th2s. It may also determine different susceptibility of Th1s and Th2s to human immunodeficiency virus strains using different fusion coreceptors.

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Differential expression of chemokine receptors in Th1s versus Th2s. 10 μg of total mRNA purified from Th1s and Th2s were used  in Northern blots analysis. Autoradiographies were obtained after 12 h of  exposure, except for CCR3 which required 7 d. Lane to lane variation in  RNA loading was <15%, as assessed by densitometric analysis of β-actin  expression. Results are representative of three experiments. (A) CCR1  through 5. (B) CXCR1 through 4.
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Figure 2: Differential expression of chemokine receptors in Th1s versus Th2s. 10 μg of total mRNA purified from Th1s and Th2s were used in Northern blots analysis. Autoradiographies were obtained after 12 h of exposure, except for CCR3 which required 7 d. Lane to lane variation in RNA loading was <15%, as assessed by densitometric analysis of β-actin expression. Results are representative of three experiments. (A) CCR1 through 5. (B) CXCR1 through 4.

Mentions: We generated Th1 and Th2 lines by stimulating human cord blood lymphocytes with mitogen in the presence of IL-12 and neutralizing anti–IL-4 mAb or IL-4 and neutralizing anti–IL-12 mAb, respectively. As previously reported, this protocol allows the establishment of human T cell lines with strongly polarized cytokine production (8). As shown at the single cell level by measuring intracellular cytokine production (Fig. 1), neonatal T cells primed under the Th1 conditions differentiated into T cells producing IFN-γ but little IL-4, whereas naive T cells primed in the presence of IL-4 and anti–IL-12 resulted in a population of T cells producing mainly IL-4. To assess the selective expression of chemokine receptors in the different T cell subsets, we have analyzed the messenger RNA (mRNA) expression levels for the different chemokine receptors in both Th1 and Th2 lines, as well as in naive cells. As shown in Fig. 2, polarized Th1s and Th2s showed differential expression of chemokine receptors. Naive T cells had a very low level of all chemokine receptor mRNAs examined except for CXCR4, the receptor for stromal cell–derived factor 1 (SDF-1; data not shown). This finding is consistent with the view that SDF-1, constitutively expressed in a broad range of tissues, is involved in basal trafficking of naive lymphocytes (27). The receptor for MCP-1 through 4 (CCR2) was expressed at high levels in both Th1s and Th2s, with somewhat higher levels for Th1s and considerable variation among different cell preparations. CCR1 (regulated on activation, normal T cell expressed and secreted [RANTES], MIP-1α, MCP-3 receptor) was equally expressed in Th1s and Th2s. In contrast, CCR4 (thymus- and activation-regulated chemokine [TARC] and MDC receptor; references 19, 28) was expressed at much higher levels in Th2s versus Th1s (14.6-fold difference by densitometry). CCR3 (eotaxin and MCP-3 receptor) was expressed only in Th2s, though at very low level in bulk cultures, which required extremely long (7 d) exposure of the blots. In contrast, the MIP-1β receptor CCR5 was preferentially (4.9-fold) expressed in Th1s versus Th2s. When CXC chemokine receptors were studied, Th1 and Th2 had no detectable CXCR1 and CXCR2 (IL-8 receptors) and equal amounts of CXCR4 (SDF-1 receptor). Moreover, expression of CXCR3, the receptor for IP-10, I-TAC, and Mig (29) was selective for Th1s (6.7-fold difference). The results described are representative of four different preparations of Th1 and Th2 lines. The preferential expression of CXCR3 and CCR5 in Th1s and CCR4 in Th2s was further confirmed on two fully differentiated Th1 (ET3.22 and ET3.20) and two Th2 (E4.1 and D4.11) clones specific for the Lol p1 antigen, and on two Th1 clones (GL93 and AC29) specific for the hepatitis delta antigen (Fig. 3 and data not shown).


Differential expression of chemokine receptors and chemotactic responsiveness of type 1 T helper cells (Th1s) and Th2s.

Bonecchi R, Bianchi G, Bordignon PP, D'Ambrosio D, Lang R, Borsatti A, Sozzani S, Allavena P, Gray PA, Mantovani A, Sinigaglia F - J. Exp. Med. (1998)

Differential expression of chemokine receptors in Th1s versus Th2s. 10 μg of total mRNA purified from Th1s and Th2s were used  in Northern blots analysis. Autoradiographies were obtained after 12 h of  exposure, except for CCR3 which required 7 d. Lane to lane variation in  RNA loading was <15%, as assessed by densitometric analysis of β-actin  expression. Results are representative of three experiments. (A) CCR1  through 5. (B) CXCR1 through 4.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2199181&req=5

Figure 2: Differential expression of chemokine receptors in Th1s versus Th2s. 10 μg of total mRNA purified from Th1s and Th2s were used in Northern blots analysis. Autoradiographies were obtained after 12 h of exposure, except for CCR3 which required 7 d. Lane to lane variation in RNA loading was <15%, as assessed by densitometric analysis of β-actin expression. Results are representative of three experiments. (A) CCR1 through 5. (B) CXCR1 through 4.
Mentions: We generated Th1 and Th2 lines by stimulating human cord blood lymphocytes with mitogen in the presence of IL-12 and neutralizing anti–IL-4 mAb or IL-4 and neutralizing anti–IL-12 mAb, respectively. As previously reported, this protocol allows the establishment of human T cell lines with strongly polarized cytokine production (8). As shown at the single cell level by measuring intracellular cytokine production (Fig. 1), neonatal T cells primed under the Th1 conditions differentiated into T cells producing IFN-γ but little IL-4, whereas naive T cells primed in the presence of IL-4 and anti–IL-12 resulted in a population of T cells producing mainly IL-4. To assess the selective expression of chemokine receptors in the different T cell subsets, we have analyzed the messenger RNA (mRNA) expression levels for the different chemokine receptors in both Th1 and Th2 lines, as well as in naive cells. As shown in Fig. 2, polarized Th1s and Th2s showed differential expression of chemokine receptors. Naive T cells had a very low level of all chemokine receptor mRNAs examined except for CXCR4, the receptor for stromal cell–derived factor 1 (SDF-1; data not shown). This finding is consistent with the view that SDF-1, constitutively expressed in a broad range of tissues, is involved in basal trafficking of naive lymphocytes (27). The receptor for MCP-1 through 4 (CCR2) was expressed at high levels in both Th1s and Th2s, with somewhat higher levels for Th1s and considerable variation among different cell preparations. CCR1 (regulated on activation, normal T cell expressed and secreted [RANTES], MIP-1α, MCP-3 receptor) was equally expressed in Th1s and Th2s. In contrast, CCR4 (thymus- and activation-regulated chemokine [TARC] and MDC receptor; references 19, 28) was expressed at much higher levels in Th2s versus Th1s (14.6-fold difference by densitometry). CCR3 (eotaxin and MCP-3 receptor) was expressed only in Th2s, though at very low level in bulk cultures, which required extremely long (7 d) exposure of the blots. In contrast, the MIP-1β receptor CCR5 was preferentially (4.9-fold) expressed in Th1s versus Th2s. When CXC chemokine receptors were studied, Th1 and Th2 had no detectable CXCR1 and CXCR2 (IL-8 receptors) and equal amounts of CXCR4 (SDF-1 receptor). Moreover, expression of CXCR3, the receptor for IP-10, I-TAC, and Mig (29) was selective for Th1s (6.7-fold difference). The results described are representative of four different preparations of Th1 and Th2 lines. The preferential expression of CXCR3 and CCR5 in Th1s and CCR4 in Th2s was further confirmed on two fully differentiated Th1 (ET3.22 and ET3.20) and two Th2 (E4.1 and D4.11) clones specific for the Lol p1 antigen, and on two Th1 clones (GL93 and AC29) specific for the hepatitis delta antigen (Fig. 3 and data not shown).

Bottom Line: In agreement with the differential chemokine receptor expression, Th1s and Th2s selectively migrate in response to the corresponding chemokines.The differential expression of chemokine receptors may dictate, to a large extent, the migration and tissue homing of Th1s and Th2s.It may also determine different susceptibility of Th1s and Th2s to human immunodeficiency virus strains using different fusion coreceptors.

View Article: PubMed Central - PubMed

Affiliation: Istituto di Ricerche Farmacologiche "Mario Negri", I-20157 Milan, Italy.

ABSTRACT
T helper cells type 1 (Th1s) that produce interferon-gamma predominantly mediate cellular immune responses and are involved in the development of chronic inflammatory conditions, whereas Th2s which produce large amounts of IL-4 and IL-5 upregulate IgE production and are prominent in the pathogenesis of allergic diseases. The precise factors determining whether Th1- or Th2-mediated immune responses preferentially occur at a peripheral site of antigen exposure are largely unknown. Chemokines, a superfamily of polypeptide mediators, are a key component of the leukocyte recruitment process. Here we report that among four CXC (CXCR1-4) and five CC (CCR1-5) chemokine receptors analyzed, CXCR3 and CCR5 are preferentially expressed in human Th1s. In contrast, Th2s preferentially express CCR4 and, to a lesser extent, CCR3. In agreement with the differential chemokine receptor expression, Th1s and Th2s selectively migrate in response to the corresponding chemokines. The differential expression of chemokine receptors may dictate, to a large extent, the migration and tissue homing of Th1s and Th2s. It may also determine different susceptibility of Th1s and Th2s to human immunodeficiency virus strains using different fusion coreceptors.

Show MeSH
Related in: MedlinePlus