Figure 6
Figure 6. Lymphangiogenesis in LNs that drain inflamed ears occurs in the absence of nodal B cells. A DTH response to oxazolone was induced in the ears of B cell-deficient JHT mice and maintained by repeatedly challenging the ears of the mice with oxazolone over 9 days. Ears and draining auricular LNs were analyzed on study day 9. (A-C) Quantitative FACS analysis detected a significant increase in the numbers of leukocytes (A) and of LECs (B) and BECs (C) in cell suspensions of inflamed compared with control (ctr) ears. (D) Immunofluorescence analysis of LYVE-1 (green) and MECA-32 (red) expression confirmed that vascularization was increased in inflamed compared with control ears. Scale bars represent 50 μm. (E,F) Analysis of ear draining auricular LNs revealed that LN weight (E) and cellularity (F) was significantly increased in LNs of inflamed animals compared with LNs of control animals. (G,H) Quantitative FACS analysis detected elevated numbers of LECs (G) in LNs draining inflamed compared with LNs draining control ears, but no change in BEC numbers (H). (I) Immunofluorescence analysis of LYVE-1 (green) and MECA-32 (red) expression confirmed that lymphatic structures were markedly expanded in LNs draining inflamed compared with control ears. Scale bars represent 100 μm. **P < .01; ***P < .001 (compared with control). Error bars are SE.

Lymphangiogenesis in LNs that drain inflamed ears occurs in the absence of nodal B cells. A DTH response to oxazolone was induced in the ears of B cell-deficient JHT mice and maintained by repeatedly challenging the ears of the mice with oxazolone over 9 days. Ears and draining auricular LNs were analyzed on study day 9. (A-C) Quantitative FACS analysis detected a significant increase in the numbers of leukocytes (A) and of LECs (B) and BECs (C) in cell suspensions of inflamed compared with control (ctr) ears. (D) Immunofluorescence analysis of LYVE-1 (green) and MECA-32 (red) expression confirmed that vascularization was increased in inflamed compared with control ears. Scale bars represent 50 μm. (E,F) Analysis of ear draining auricular LNs revealed that LN weight (E) and cellularity (F) was significantly increased in LNs of inflamed animals compared with LNs of control animals. (G,H) Quantitative FACS analysis detected elevated numbers of LECs (G) in LNs draining inflamed compared with LNs draining control ears, but no change in BEC numbers (H). (I) Immunofluorescence analysis of LYVE-1 (green) and MECA-32 (red) expression confirmed that lymphatic structures were markedly expanded in LNs draining inflamed compared with control ears. Scale bars represent 100 μm. **P < .01; ***P < .001 (compared with control). Error bars are SE.

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