Isolation of the intact white pulp. Quantitative and qualitative analysis of the cellular composition of the splenic compartments

Abstract

The spleen is anatomically and functionally divided into two compartments: the red pulp, where particles are effectively removed from the blood, and the white pulp, where specific immune responses are generated. Here the isolation of white pulp from red pulp is described, allowing a detailed analysis of the cellular components of both red and white pulp separately. A striking abundance of memory T cells was found in the white and red pulp with an overall ratio of T and B cells in the white pulp being similar to that in lymph nodes. Both NK and gamma delta T cells can be found in white pulp and lymph nodes, but granulocytes are absent. The distribution of dendritic cell subsets showed significant differences between white pulp and lymph nodes. Furthermore, short-term homing experiments showed that migration of lymphocytes into the white pulp greatly exceeded that into lymph nodes, with significant differences in migration of various lymphocytes subsets. This suggests a different migration and retention mechanism in the white pulp. This new isolation technique will allow further analysis of the functional capacities of the splenic compartments.

Ageing adversely affects the migration and function of marginal zone B cells

“…Binding of anti‐CD21‐PE mAb by some follicular B cells was also evident at this time (Fig. 3b), consistent with the presence of recirculating follicular B cells in the MZ and blood 3, 23. By 20 min after anti‐CD21‐PE mAb injection the frequency of PE + MZ B cells had increased to c. 80% in young mice (Fig.…”

Blocking L-selectin and α4-integrin changes donor cell homing pattern and ameliorates murine acute graft versus host disease

“…It has long been known that lymphocytes employ different mechanisms in homing into LN and the spleen [20][21][22][23][24]. While the factors that influence lymphocyte homing into the spleen have not been understood, those affecting the homing into LN have been extensively studied and partially elucidated, especially using gene knockout mice [20][21][22][23][24]. Circulating lymphocytes must first adhere to microvascular endothelial cells in the presence of hydrodynamic shear exerted by the flowing blood.…”

“…Ab directed at L-selectin (CD62L) and § 4 integrins (CD49d) were only partially effective in blocking the homing of donor cells into the recipient LN, probably in part due to an in vivo dissociation of the Ab from the cells. While the mAb that directed at CD62L failed to reduce the homing of donor cells into the spleen of the recipient mice, anti-CD49d significantly enhanced this homing process, indicating again that lymphocytes use a different approach in homing into LN and spleens [20][21][22][23][24].…”

The selective expression of distinct fucosylated glycoproteins on murine T and B lymphocyte subsets

“…In adult mice, however, 13% and 9% of the total splenocytes were positively labeled with UEA-I and AAA, respectively. Bearing in mind the phenotypic heterogeneity of the discrete lymphocyte subsets inhabiting the splenic compartments and the possible age-dependent modulation of their migration patterns (Timens et al, 1989;Butcher et al, 1999;Loder et al, 1999;Nolte et al, 2000;Lo et al, 2003), the identity of the UEA-I + and AAA + lymphocytes in both age groups of mice was further gauged in labeling assays conducted with isolated splenic lymphocyte subpopulations.…”