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Lymphocyte migration in lymphocyte function-associated antigen (LFA)-1-deficient mice.

Berlin-Rufenach C, Otto F, Mathies M, Westermann J, Owen MJ, Hamann A, Hogg N - J. Exp. Med. (1999)

Bottom Line: LFA-1 has a key role in migration to peripheral lymph nodes (pLNs), and influences migration into other LNs.Second, the alpha4 integrins, alpha4beta7 and alpha4beta1, have a hitherto unrecognized ability to compensate for the lack of LFA-1 in migration to pLNs.VCAM-1 also participates in trafficking into mesenteric LNs and Peyer's patch nodes where mucosal addressin cell adhesion molecule 1 (MAdCAM-1), the alpha4beta7-specific ligand, dominates.

View Article: PubMed Central - PubMed

Affiliation: Leukocyte Adhesion Laboratory, Imperial Cancer Research Fund, London WC2A 3PX, United Kingdom.

ABSTRACT
Using lymphocyte function-associated antigen (LFA)-1(-/-) mice, we have examined the role of LFA-1 and other integrins in the recirculation of lymphocytes. LFA-1 has a key role in migration to peripheral lymph nodes (pLNs), and influences migration into other LNs. Second, the alpha4 integrins, alpha4beta7 and alpha4beta1, have a hitherto unrecognized ability to compensate for the lack of LFA-1 in migration to pLNs. These findings are confirmed using normal mice and blocking LFA-1 and alpha4 monoclonal antibodies. Unexpectedly, vascular cell adhesion molecule (VCAM)-1, which is essential in inflammatory responses, serves as the ligand for the alpha4 integrins on pLN high endothelial venules. VCAM-1 also participates in trafficking into mesenteric LNs and Peyer's patch nodes where mucosal addressin cell adhesion molecule 1 (MAdCAM-1), the alpha4beta7-specific ligand, dominates. Both alpha4beta1, interacting with ligand VCAM-1, and also LFA-1 participate in substantial lymphocyte recirculation through bone marrow. These observations suggest that organ-specific adhesion receptor usage in mature lymphocyte recirculation is not as rigidly adhered to as previously considered, and that the same basic sets of adhesion receptors are used in both lymphocyte homing and inflammatory responses.

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Differential rates of homing of CT-labeled lymphocytes from  wild-type and LFA-1−/− mice. (a) Two-color FACS® analysis of the cell  mixture of LFA-1−/− and LFA-1+/+ cells injected intravenously into a wild-type C57BL/6 mouse (left) and the fluorescent cells recovered from the  pLNs after a 1-h homing period (right). For details, see Materials and Methods. (b) Homing experiments show that within 1 h, LFA-1−/− lymphocytes  migrate into pLNs, mLNs, and PPs with 21, 51, and 68% of the efficiency of  LFA-1+/+ lymphocytes, respectively. Conversely, LFA-1–deficient lymphocytes occur in larger numbers in the spleen than do wild-type cells (ratio of  1.29). These data are averaged from 10–12 experiments. In all cases, the  mean ratios are significantly different from 1.0 (P < 0.05) as assessed by t test.
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Figure 3: Differential rates of homing of CT-labeled lymphocytes from wild-type and LFA-1−/− mice. (a) Two-color FACS® analysis of the cell mixture of LFA-1−/− and LFA-1+/+ cells injected intravenously into a wild-type C57BL/6 mouse (left) and the fluorescent cells recovered from the pLNs after a 1-h homing period (right). For details, see Materials and Methods. (b) Homing experiments show that within 1 h, LFA-1−/− lymphocytes migrate into pLNs, mLNs, and PPs with 21, 51, and 68% of the efficiency of LFA-1+/+ lymphocytes, respectively. Conversely, LFA-1–deficient lymphocytes occur in larger numbers in the spleen than do wild-type cells (ratio of 1.29). These data are averaged from 10–12 experiments. In all cases, the mean ratios are significantly different from 1.0 (P < 0.05) as assessed by t test.

Mentions: To gain further information about the diminished cellularity of the pLNs and to investigate trafficking capabilities of LFA-1–deficient lymphocytes, short-term migration studies were performed. The approach taken was to label LFA-1−/− and LFA-1+/+ lymphocytes with two distinguishable CT orange and green fluorescent dyes and inject equivalent numbers into the tail vein of C57BL/6 recipients. This allowed direct comparison of the homing activity of the lymphocytes within each LN setting (29). An example of the methodology is illustrated in Fig. 3 a. When the pLN lymphocytes were examined for proportions of CT orange to green after 1 h of homing, it was observed that ∼13% of LFA-1−/− cells compared with LFA-1+/+ cells had migrated into the pLNs (Fig. 3 a). We then examined the relative ability of LFA-1−/− cells to gain entry into other secondary lymphoid tissues (Fig. 3 b). When compared with LFA-1+/+, the LFA-1−/− cells were most poorly represented in pLNs (0.21 ± 0.01), followed by mLNs (0.51 ± 0.02) and PPs (0.68 ± 0.03). In contrast, there was a marked increase in the LFA-1−/− cells in the spleen (1.29 ± 0.04) which is likely due to redistribution of lymphocytes from LN to spleen. These findings on the effect of LFA-1 absence on the trafficking of lymphocytes were confirmed using the technique of tracking 51Cr-labeled lymphocytes to compare homing in LFA-1−/− and LFA-1+/+ mice (data not shown). When CD4, CD8, and B220+ lymphocyte subsets were analyzed separately, the ratio of migrated to injected lymphocytes was identical among the subsets whether derived from wild-type or LFA-1−/− mice, indicating that the lack of LFA-1 caused equal difficulty for all types of lymphocytes to gain entry into the LNs (data not shown).


Lymphocyte migration in lymphocyte function-associated antigen (LFA)-1-deficient mice.

Berlin-Rufenach C, Otto F, Mathies M, Westermann J, Owen MJ, Hamann A, Hogg N - J. Exp. Med. (1999)

Differential rates of homing of CT-labeled lymphocytes from  wild-type and LFA-1−/− mice. (a) Two-color FACS® analysis of the cell  mixture of LFA-1−/− and LFA-1+/+ cells injected intravenously into a wild-type C57BL/6 mouse (left) and the fluorescent cells recovered from the  pLNs after a 1-h homing period (right). For details, see Materials and Methods. (b) Homing experiments show that within 1 h, LFA-1−/− lymphocytes  migrate into pLNs, mLNs, and PPs with 21, 51, and 68% of the efficiency of  LFA-1+/+ lymphocytes, respectively. Conversely, LFA-1–deficient lymphocytes occur in larger numbers in the spleen than do wild-type cells (ratio of  1.29). These data are averaged from 10–12 experiments. In all cases, the  mean ratios are significantly different from 1.0 (P < 0.05) as assessed by t test.
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Related In: Results  -  Collection

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Figure 3: Differential rates of homing of CT-labeled lymphocytes from wild-type and LFA-1−/− mice. (a) Two-color FACS® analysis of the cell mixture of LFA-1−/− and LFA-1+/+ cells injected intravenously into a wild-type C57BL/6 mouse (left) and the fluorescent cells recovered from the pLNs after a 1-h homing period (right). For details, see Materials and Methods. (b) Homing experiments show that within 1 h, LFA-1−/− lymphocytes migrate into pLNs, mLNs, and PPs with 21, 51, and 68% of the efficiency of LFA-1+/+ lymphocytes, respectively. Conversely, LFA-1–deficient lymphocytes occur in larger numbers in the spleen than do wild-type cells (ratio of 1.29). These data are averaged from 10–12 experiments. In all cases, the mean ratios are significantly different from 1.0 (P < 0.05) as assessed by t test.
Mentions: To gain further information about the diminished cellularity of the pLNs and to investigate trafficking capabilities of LFA-1–deficient lymphocytes, short-term migration studies were performed. The approach taken was to label LFA-1−/− and LFA-1+/+ lymphocytes with two distinguishable CT orange and green fluorescent dyes and inject equivalent numbers into the tail vein of C57BL/6 recipients. This allowed direct comparison of the homing activity of the lymphocytes within each LN setting (29). An example of the methodology is illustrated in Fig. 3 a. When the pLN lymphocytes were examined for proportions of CT orange to green after 1 h of homing, it was observed that ∼13% of LFA-1−/− cells compared with LFA-1+/+ cells had migrated into the pLNs (Fig. 3 a). We then examined the relative ability of LFA-1−/− cells to gain entry into other secondary lymphoid tissues (Fig. 3 b). When compared with LFA-1+/+, the LFA-1−/− cells were most poorly represented in pLNs (0.21 ± 0.01), followed by mLNs (0.51 ± 0.02) and PPs (0.68 ± 0.03). In contrast, there was a marked increase in the LFA-1−/− cells in the spleen (1.29 ± 0.04) which is likely due to redistribution of lymphocytes from LN to spleen. These findings on the effect of LFA-1 absence on the trafficking of lymphocytes were confirmed using the technique of tracking 51Cr-labeled lymphocytes to compare homing in LFA-1−/− and LFA-1+/+ mice (data not shown). When CD4, CD8, and B220+ lymphocyte subsets were analyzed separately, the ratio of migrated to injected lymphocytes was identical among the subsets whether derived from wild-type or LFA-1−/− mice, indicating that the lack of LFA-1 caused equal difficulty for all types of lymphocytes to gain entry into the LNs (data not shown).

Bottom Line: LFA-1 has a key role in migration to peripheral lymph nodes (pLNs), and influences migration into other LNs.Second, the alpha4 integrins, alpha4beta7 and alpha4beta1, have a hitherto unrecognized ability to compensate for the lack of LFA-1 in migration to pLNs.VCAM-1 also participates in trafficking into mesenteric LNs and Peyer's patch nodes where mucosal addressin cell adhesion molecule 1 (MAdCAM-1), the alpha4beta7-specific ligand, dominates.

View Article: PubMed Central - PubMed

Affiliation: Leukocyte Adhesion Laboratory, Imperial Cancer Research Fund, London WC2A 3PX, United Kingdom.

ABSTRACT
Using lymphocyte function-associated antigen (LFA)-1(-/-) mice, we have examined the role of LFA-1 and other integrins in the recirculation of lymphocytes. LFA-1 has a key role in migration to peripheral lymph nodes (pLNs), and influences migration into other LNs. Second, the alpha4 integrins, alpha4beta7 and alpha4beta1, have a hitherto unrecognized ability to compensate for the lack of LFA-1 in migration to pLNs. These findings are confirmed using normal mice and blocking LFA-1 and alpha4 monoclonal antibodies. Unexpectedly, vascular cell adhesion molecule (VCAM)-1, which is essential in inflammatory responses, serves as the ligand for the alpha4 integrins on pLN high endothelial venules. VCAM-1 also participates in trafficking into mesenteric LNs and Peyer's patch nodes where mucosal addressin cell adhesion molecule 1 (MAdCAM-1), the alpha4beta7-specific ligand, dominates. Both alpha4beta1, interacting with ligand VCAM-1, and also LFA-1 participate in substantial lymphocyte recirculation through bone marrow. These observations suggest that organ-specific adhesion receptor usage in mature lymphocyte recirculation is not as rigidly adhered to as previously considered, and that the same basic sets of adhesion receptors are used in both lymphocyte homing and inflammatory responses.

Show MeSH
Related in: MedlinePlus