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Rapid acquisition of tissue-specific homing phenotypes by CD4(+) T cells activated in cutaneous or mucosal lymphoid tissues.

Campbell DJ, Butcher EC - J. Exp. Med. (2002)

Bottom Line: However, little is known about the development of these selectively homing T cell subsets, and it is unclear whether activation in cutaneous versus intestinal lymphoid organs directly results in effector/memory T cells that differentially express adhesion and chemoattractant receptors targeting them to the corresponding nonlymphoid site.We define two murine CD4(+) effector/memory T cell subsets that preferentially localize in cutaneous or intestinal lymphoid organs by their reciprocal expression of the adhesion molecules P-selectin ligand (P-lig) and alpha 4 beta 7, respectively.Thus, during an immune response, local microenvironments within cutaneous and intestinal secondary lymphoid organs differentially direct T cell expression of these adhesion and chemoattractant receptors, targeting the resulting effector T cells to the inflamed skin or intestinal lamina propria.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Immunology and Vascular Biology, Department of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA. daniel@macampbell.com

ABSTRACT
Effector and memory T cells can be subdivided based on their ability to traffic through peripheral tissues such as inflamed skin and intestinal lamina propria, a property controlled by expression of 'tissue-specific' adhesion and chemoattractant receptors. However, little is known about the development of these selectively homing T cell subsets, and it is unclear whether activation in cutaneous versus intestinal lymphoid organs directly results in effector/memory T cells that differentially express adhesion and chemoattractant receptors targeting them to the corresponding nonlymphoid site. We define two murine CD4(+) effector/memory T cell subsets that preferentially localize in cutaneous or intestinal lymphoid organs by their reciprocal expression of the adhesion molecules P-selectin ligand (P-lig) and alpha 4 beta 7, respectively. We show that within 2 d of systemic immunization CD4(+) T cells activated in cutaneous lymph nodes upregulate P-lig, and downregulate alpha 4 beta 7, while those responding to antigen in intestinal lymph nodes selectively express high levels of alpha 4 beta 7 and acquire responsiveness to the intestinal chemokine thymus-expressed chemokine (TECK). Thus, during an immune response, local microenvironments within cutaneous and intestinal secondary lymphoid organs differentially direct T cell expression of these adhesion and chemoattractant receptors, targeting the resulting effector T cells to the inflamed skin or intestinal lamina propria.

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CD4+ T cells activated in PLNs and MLNs differentially upregulate P-lig and α4β7. (A) Percentage of OVA-specific KJ1–26+ cells among gated CD4+ T cells isolated from peripheral blood at the indicated times after intraperitoneal injection of OVA plus LPS. Data are mean and SD of values obtained from five mice at each time point. (B) Representative flow cytometry data of cellular CFSE content and KJ1–26 staining on gated CD4+ T cells isolated from the indicated tissues 2 d after intraperitoneal immunization of DO11.10 adoptive transfer recipients with OVA plus LPS. The horizontal marker in the histograms represents the CFSE fluorescence intensity of naive CD4+KJ1–26+ T cells isolated from animals immunized with LPS alone (data not shown). (C) Expression of α4β7 and P-lig by gated CD4+KJ1–26+ cells isolated from PLNs (• and ○) and MLNs (▪ and □) 2 d after intraperitoneal immunization of DO11.10 adoptive transfer recipients with OVA plus LPS (black symbols) or LPS alone (white symbols). Each data point represents a measurement from an individual animal. (D) Representative flow cytometry data of α4β7 and P-lig staining on gated CD4+KJ1–26+ cells isolated from PLNs (top) and MLN (bottom) 2 d after immunization of DO11.10 adoptive transfer recipients with OVA plus LPS (left) or LPS alone (right). The quadrant gate used to define the P-lig+ and α4β7hi populations in C is indicated. (E) Mean fluorescence intensity (MFI) of α4β7 (left) and P-lig (right) staining on gated CD4+KJ1–26+ cells isolated from the MLNs (▪) or PLNs (•) 2 d after intraperitoneal immunization of DO11.10 adoptive transfer recipients with OVA plus LPS as a function of cell division (as determined by CFSE content). Data are mean and SE of values obtained from four (α4β7) or five (P-lig) mice. N represents the MFI of α4β7 or P-lig staining on naive cells isolated from animals immunized with LPS alone. Dotted lines represent background MFI of cells stained with an isotype control (left) or unstained cells (right).
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fig3: CD4+ T cells activated in PLNs and MLNs differentially upregulate P-lig and α4β7. (A) Percentage of OVA-specific KJ1–26+ cells among gated CD4+ T cells isolated from peripheral blood at the indicated times after intraperitoneal injection of OVA plus LPS. Data are mean and SD of values obtained from five mice at each time point. (B) Representative flow cytometry data of cellular CFSE content and KJ1–26 staining on gated CD4+ T cells isolated from the indicated tissues 2 d after intraperitoneal immunization of DO11.10 adoptive transfer recipients with OVA plus LPS. The horizontal marker in the histograms represents the CFSE fluorescence intensity of naive CD4+KJ1–26+ T cells isolated from animals immunized with LPS alone (data not shown). (C) Expression of α4β7 and P-lig by gated CD4+KJ1–26+ cells isolated from PLNs (• and ○) and MLNs (▪ and □) 2 d after intraperitoneal immunization of DO11.10 adoptive transfer recipients with OVA plus LPS (black symbols) or LPS alone (white symbols). Each data point represents a measurement from an individual animal. (D) Representative flow cytometry data of α4β7 and P-lig staining on gated CD4+KJ1–26+ cells isolated from PLNs (top) and MLN (bottom) 2 d after immunization of DO11.10 adoptive transfer recipients with OVA plus LPS (left) or LPS alone (right). The quadrant gate used to define the P-lig+ and α4β7hi populations in C is indicated. (E) Mean fluorescence intensity (MFI) of α4β7 (left) and P-lig (right) staining on gated CD4+KJ1–26+ cells isolated from the MLNs (▪) or PLNs (•) 2 d after intraperitoneal immunization of DO11.10 adoptive transfer recipients with OVA plus LPS as a function of cell division (as determined by CFSE content). Data are mean and SE of values obtained from four (α4β7) or five (P-lig) mice. N represents the MFI of α4β7 or P-lig staining on naive cells isolated from animals immunized with LPS alone. Dotted lines represent background MFI of cells stained with an isotype control (left) or unstained cells (right).

Mentions: Adoptive transfer recipients were immunized with a single intraperitoneal injection of OVA+LPS. LPS was chosen as an adjuvant for its known proinflammatory effects and its ability to potentiate immune responses against soluble proteins. Consistent with the ‘antigen-trapping’ of T cells in secondary lymphoid tissues (15), the antigen-specific KJ1–26+ T cells disappeared from the recirculating pool of T cells in the blood within 1 d of immunization, where they were not detected again in significant numbers until day 3 (Fig. 3 A). Cell division analysis using CFSE-labeled transgenic cells reveals that during this period of sequestration from the blood, antigen-specific T cells were activated and proliferating in both subcutaneous PLNs and MLNs, where by day 2 after immunization most cells in both tissues had divided ∼2–4 times (Fig. 3 B). Thus, this adoptive transfer and immunization protocol provides a unique opportunity to compare the surface phenotype of antigen-specific CD4+ T cells activated in the PLNs or MLNs, before these populations have intermingled through effector cell recirculation. Accordingly, we analyzed expression of P-lig and α4β7 on antigen-specific T cells 2 d after immunization during this period of sequestration in the lymph nodes. While cells isolated from animals immunized with LPS alone maintained their naive phenotype, we found that even at this very early time point a major fraction of the cells activated in the MLNs expressed high levels of α4β7 (30–60%), whereas P-lig was preferentially expressed by a similar proportion of cells activated in subcutaneous PLNs (Fig. 3 C and D). CFSE analysis was also used to examine α4β7 integrin and P-lig expression as a function of cell division (Fig. 3 E). Cells that had undergone the same number of divisions in the PLNs or MLNs still displayed differential expression of P-lig and α4β7, and the distinct homing phenotypes of these cells cannot therefore be accounted for by subtle differences in the extent of T cell activation and proliferation in these tissues.


Rapid acquisition of tissue-specific homing phenotypes by CD4(+) T cells activated in cutaneous or mucosal lymphoid tissues.

Campbell DJ, Butcher EC - J. Exp. Med. (2002)

CD4+ T cells activated in PLNs and MLNs differentially upregulate P-lig and α4β7. (A) Percentage of OVA-specific KJ1–26+ cells among gated CD4+ T cells isolated from peripheral blood at the indicated times after intraperitoneal injection of OVA plus LPS. Data are mean and SD of values obtained from five mice at each time point. (B) Representative flow cytometry data of cellular CFSE content and KJ1–26 staining on gated CD4+ T cells isolated from the indicated tissues 2 d after intraperitoneal immunization of DO11.10 adoptive transfer recipients with OVA plus LPS. The horizontal marker in the histograms represents the CFSE fluorescence intensity of naive CD4+KJ1–26+ T cells isolated from animals immunized with LPS alone (data not shown). (C) Expression of α4β7 and P-lig by gated CD4+KJ1–26+ cells isolated from PLNs (• and ○) and MLNs (▪ and □) 2 d after intraperitoneal immunization of DO11.10 adoptive transfer recipients with OVA plus LPS (black symbols) or LPS alone (white symbols). Each data point represents a measurement from an individual animal. (D) Representative flow cytometry data of α4β7 and P-lig staining on gated CD4+KJ1–26+ cells isolated from PLNs (top) and MLN (bottom) 2 d after immunization of DO11.10 adoptive transfer recipients with OVA plus LPS (left) or LPS alone (right). The quadrant gate used to define the P-lig+ and α4β7hi populations in C is indicated. (E) Mean fluorescence intensity (MFI) of α4β7 (left) and P-lig (right) staining on gated CD4+KJ1–26+ cells isolated from the MLNs (▪) or PLNs (•) 2 d after intraperitoneal immunization of DO11.10 adoptive transfer recipients with OVA plus LPS as a function of cell division (as determined by CFSE content). Data are mean and SE of values obtained from four (α4β7) or five (P-lig) mice. N represents the MFI of α4β7 or P-lig staining on naive cells isolated from animals immunized with LPS alone. Dotted lines represent background MFI of cells stained with an isotype control (left) or unstained cells (right).
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fig3: CD4+ T cells activated in PLNs and MLNs differentially upregulate P-lig and α4β7. (A) Percentage of OVA-specific KJ1–26+ cells among gated CD4+ T cells isolated from peripheral blood at the indicated times after intraperitoneal injection of OVA plus LPS. Data are mean and SD of values obtained from five mice at each time point. (B) Representative flow cytometry data of cellular CFSE content and KJ1–26 staining on gated CD4+ T cells isolated from the indicated tissues 2 d after intraperitoneal immunization of DO11.10 adoptive transfer recipients with OVA plus LPS. The horizontal marker in the histograms represents the CFSE fluorescence intensity of naive CD4+KJ1–26+ T cells isolated from animals immunized with LPS alone (data not shown). (C) Expression of α4β7 and P-lig by gated CD4+KJ1–26+ cells isolated from PLNs (• and ○) and MLNs (▪ and □) 2 d after intraperitoneal immunization of DO11.10 adoptive transfer recipients with OVA plus LPS (black symbols) or LPS alone (white symbols). Each data point represents a measurement from an individual animal. (D) Representative flow cytometry data of α4β7 and P-lig staining on gated CD4+KJ1–26+ cells isolated from PLNs (top) and MLN (bottom) 2 d after immunization of DO11.10 adoptive transfer recipients with OVA plus LPS (left) or LPS alone (right). The quadrant gate used to define the P-lig+ and α4β7hi populations in C is indicated. (E) Mean fluorescence intensity (MFI) of α4β7 (left) and P-lig (right) staining on gated CD4+KJ1–26+ cells isolated from the MLNs (▪) or PLNs (•) 2 d after intraperitoneal immunization of DO11.10 adoptive transfer recipients with OVA plus LPS as a function of cell division (as determined by CFSE content). Data are mean and SE of values obtained from four (α4β7) or five (P-lig) mice. N represents the MFI of α4β7 or P-lig staining on naive cells isolated from animals immunized with LPS alone. Dotted lines represent background MFI of cells stained with an isotype control (left) or unstained cells (right).
Mentions: Adoptive transfer recipients were immunized with a single intraperitoneal injection of OVA+LPS. LPS was chosen as an adjuvant for its known proinflammatory effects and its ability to potentiate immune responses against soluble proteins. Consistent with the ‘antigen-trapping’ of T cells in secondary lymphoid tissues (15), the antigen-specific KJ1–26+ T cells disappeared from the recirculating pool of T cells in the blood within 1 d of immunization, where they were not detected again in significant numbers until day 3 (Fig. 3 A). Cell division analysis using CFSE-labeled transgenic cells reveals that during this period of sequestration from the blood, antigen-specific T cells were activated and proliferating in both subcutaneous PLNs and MLNs, where by day 2 after immunization most cells in both tissues had divided ∼2–4 times (Fig. 3 B). Thus, this adoptive transfer and immunization protocol provides a unique opportunity to compare the surface phenotype of antigen-specific CD4+ T cells activated in the PLNs or MLNs, before these populations have intermingled through effector cell recirculation. Accordingly, we analyzed expression of P-lig and α4β7 on antigen-specific T cells 2 d after immunization during this period of sequestration in the lymph nodes. While cells isolated from animals immunized with LPS alone maintained their naive phenotype, we found that even at this very early time point a major fraction of the cells activated in the MLNs expressed high levels of α4β7 (30–60%), whereas P-lig was preferentially expressed by a similar proportion of cells activated in subcutaneous PLNs (Fig. 3 C and D). CFSE analysis was also used to examine α4β7 integrin and P-lig expression as a function of cell division (Fig. 3 E). Cells that had undergone the same number of divisions in the PLNs or MLNs still displayed differential expression of P-lig and α4β7, and the distinct homing phenotypes of these cells cannot therefore be accounted for by subtle differences in the extent of T cell activation and proliferation in these tissues.

Bottom Line: However, little is known about the development of these selectively homing T cell subsets, and it is unclear whether activation in cutaneous versus intestinal lymphoid organs directly results in effector/memory T cells that differentially express adhesion and chemoattractant receptors targeting them to the corresponding nonlymphoid site.We define two murine CD4(+) effector/memory T cell subsets that preferentially localize in cutaneous or intestinal lymphoid organs by their reciprocal expression of the adhesion molecules P-selectin ligand (P-lig) and alpha 4 beta 7, respectively.Thus, during an immune response, local microenvironments within cutaneous and intestinal secondary lymphoid organs differentially direct T cell expression of these adhesion and chemoattractant receptors, targeting the resulting effector T cells to the inflamed skin or intestinal lamina propria.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Immunology and Vascular Biology, Department of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA. daniel@macampbell.com

ABSTRACT
Effector and memory T cells can be subdivided based on their ability to traffic through peripheral tissues such as inflamed skin and intestinal lamina propria, a property controlled by expression of 'tissue-specific' adhesion and chemoattractant receptors. However, little is known about the development of these selectively homing T cell subsets, and it is unclear whether activation in cutaneous versus intestinal lymphoid organs directly results in effector/memory T cells that differentially express adhesion and chemoattractant receptors targeting them to the corresponding nonlymphoid site. We define two murine CD4(+) effector/memory T cell subsets that preferentially localize in cutaneous or intestinal lymphoid organs by their reciprocal expression of the adhesion molecules P-selectin ligand (P-lig) and alpha 4 beta 7, respectively. We show that within 2 d of systemic immunization CD4(+) T cells activated in cutaneous lymph nodes upregulate P-lig, and downregulate alpha 4 beta 7, while those responding to antigen in intestinal lymph nodes selectively express high levels of alpha 4 beta 7 and acquire responsiveness to the intestinal chemokine thymus-expressed chemokine (TECK). Thus, during an immune response, local microenvironments within cutaneous and intestinal secondary lymphoid organs differentially direct T cell expression of these adhesion and chemoattractant receptors, targeting the resulting effector T cells to the inflamed skin or intestinal lamina propria.

Show MeSH
Related in: MedlinePlus