B cells isolated from germinal centers (GC) of immune mice 2-5 days after antigen (Ag) challenge migrate in response to chemotactic signals, whereas GC B cells isolated at other times and resting B cells do not. Since B cells are in direct contact with follicular dendritic cells (FDC) in GC we reasoned that FDC might play a role in enabling B cells to become chemotactically active. Resting B cells were co-cultured with FDC either with or without anti-mu-dextran (anti-mu-dex) as an Ag surrogate and/or recombinant interleukin (rIL)-4 as a T cell surrogate. After 3 days, the B cells were isolated and their migration to chemotactic factors contained in zymosan-activated serum assessed in microchemotaxis chambers. B cells incubated alone or with anti-mu-dex or rIL-4 showed minimal migration, which could be increased if both anti-mu-dex or rIL-4 were present. However, maximal migration was obtained when B cells were cultured with FDC, and this was not increased by addition of anti-mu-dex and/or rIL-4, indicating that the FDC signal was a primary signal and did not require pre-activation of the B cells. Checkerboard analysis using variation in concentration and location of the chemoattractant in chemotaxis chambers indicated that both chemotaxis and chemokinesis occurred. B cell migration began within 6 h of culture, peaked by 48 h and decreased thereafter. Removal of FDC or interference with FDC-B cell contact ablated or significantly decreased induction of B cell migration. Furthermore, induction did not require functional T cells. These data indicate that FDC can induce resting B cells to become responsive to chemotactic signals.
The handpiece of a Florida sleeve probe was modified to create a flange with the capability to detect the cemento-enamel junction (CEJ). This new instrument (the Pressure-controlled, Automated, Standardised Handpiece or Florida PASHA probe) was used to determine whether (a) the CEJ could be reproducibly detected in dried, human skulls and (b) clinical attachment levels could be reliably measured in human subjects. When using the Florida PASHA probe to detect the CEJ at 157 different sites in four dried, human skulls, there were no statistically significant (p > or = 0.15) differences in mean CEJ detection measurements for any of the three participating examiners; either when the CEJ was visualised or obscured. The mean differences between first and second replicates ranged from 0.00 to 0.08 mm. Intraclass correlation coefficients (ICCs) of repeated measures in both conditions ranged from 0.70 to 0.83 for tactile CEJ detection (CEJ obscured), and from 0.95 to 0.96 for visual detection (CEJ visible). In human studies, the Florida PASHA probe was used by two examiners to determine clinical attachment levels (CAL) at 660 sites in 5 human subjects undergoing supportive periodontal therapy. Intra-examiner agreement of replicate measurements recorded by the probe, as measured by calculating ICCs, ranged from 0.79 to 0.85 for the 2 examiners, respectively. A statistically significant inter-examiner difference in mean CAL measurements when using the Florida PASHA probe was found (p<0.001). Notwithstanding this difference, inter-examiner agreement was good, with an ICC of 0.83. These data suggest that the Florida PASHA probe can reproducibly detect the CEJ and is proposed as a tool for measuring CAL in humans.
An infiltrate of B cells and plasma cells is characteristic of certain chronic inflammatory lesions. However, mechanisms involved in the local accumulation of these cells have not been established. Efforts to demonstrate that B cells from normal animals can migrate in response to inflammation-induced chemoattractants have been inconclusive. The objective of this study was to determine if murine germinal center (GC) B cells could respond chemotactically to a C5a gradient. On successive days after secondary immunization, draining lymph nodes were harvested and the activated GC B cells isolated. These GC B cells were placed in modified Boyden chambers, incubated for 3 h and the distance the leading front of cells migrated through the filters was determined. The results show that GC B cells migrated to factors in zymosan- and lipopolysaccharide-activated serum. The migratory response demonstrated distinct kinetics. Cells isolated between 2 to 4 days after secondary immunization migrated, whereas cells isolated at day 0 and beyond day 6 did not. Checkerboard analysis revealed that the migratory response was attributable to both chemokinesis and chemotaxis. Anti-C5 inhibited the migration of day-3 GC B cells implicating C5 in the migration mechanism. Studies using recombinant C5a established that this C5 fragment was chemotactically active. In conclusion, GC B cells generally were not chemotactically active. However, at a particular stage of maturation B cells in the GC become responsive to C5a as a chemotactic agent. Thus, B cells from normal animals may respond chemotactically, and C5a may play a role in recruitment of recently activated B cells into inflammatory sites.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
customersupport@researchsolutions.com
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.