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Updated: Apr 14, 2026

Isolating And Immunostaining Lymphocytes and Dendritic Cells from Murine Peyer's Patches
Published on: March 17, 2013
[Biological roles of somatostatin in intestinal macaque Peyer's patch B cells]
Hao Wu1, Ling Liu1, Qinghua Tan1
1Department of Gastroenterology, West China Hospital, Sichuan University, Chengdu 610041, China.
Objective:
To explore the effects of somatostatin (SST) on macaque intestinal Peyer's patches (PP) in immune B cells and explore its mechanism.
Methods:
A total of 15 healthy adult macaques were randomized into control,MODS and MODS+SST groups.Surgical procedures of MODS in macaques: For MODS group, anesthesia was maintained with diazepam (0.16 ± 0.09) mg×kg(-1)×h(-1), i.v.). A catheter was inserted into a peripheral vein for infusing 0.9% saline and 20 g glucose (0.1-0.2 ml×kg(-1)×min(-1), i.v.gtt) for 24 h. Midline laparotomy was performed. Then superior mesenteric artery (SMA) was isolated and occluded with a microsurgical clip. After a 1-hour occlusion, clip was removed and intestinal perfusion reestablished.In MODS+SST group, SST was infused intravenously with a syringe pump at a dosage of 5 µg×kg(-1)×h(-1) for 5 min before an occlusion of SMA until the end of experiment.Venous blood samples were redrawn and the animals sacrificed at 24 h post-IIR for harvesting vital organs. The changes of organs and the morphological changes of PP were detected by hematoxylin and eosin. And the expressions of TLR4, TLR2, CD20, CD5, α4β7 and MadCAM-1 were evaluated by immunohistochemical staining.Semiquantitative immunohistochemical analysis of raw data was performed with Image Pro Plus 4.0 software.
Results:
All animals in MODS group presented with small intestines PP increased both in number and size compared with control group (4.8 ± 2.3 vs 1.2 ± 0.9, P < 0.05). After prophylactic to SST, compared with MODS group, the number of PP in small intestines in MODS+SST group decreased (2.7 ± 1.5 vs 4.8 ± 2.3, P < 0.05), but there was no significant difference in morphological of PP. The expression of CD20(+) of B-cells in MODS group was significantly lower than in normal group (integrated optical density (IOD), 64.22 ± 42.45 vs 100.00 ± 86.67, P < 0.05).Interestingly, after prophylactic dosing of SST, the expression level of CD20(+) of B-cells elevated significantly in MODS+SST group compared with that in MODS group (129.02 ± 75.04 vs 64.22 ± 42.45, P < 0.05). There was no α4β7 expression of B-cells in PP of macaques in three groups. And the expression level of MadCAM-1 was strongly up-regulated after prophylactic dosing of STT compared with MODS. After prophylactic dosing of STT, the expressions of TLR4 and TLR2 declined significantly compared with MODS (56.60 ± 6.83 vs 93.26 ± 10.40, 35.56 ± 4.71 vs 62.06 ± 9.90, P < 0.05). Plasma cells within normal intestinal mucosa were predominantly located in lamina propria. During MODS, plasma cells almost disappeared. After prophylactic dosing of STT, compared to MODS group, there was a rebound of plasma cells.
Conclusions:
Endogenous SST controls the switch of B-cells in PP between natural and acquired immunities. The prophylactic dosing of SST could convert excessive natural immunity into beneficial acquired humoral immunity. And SST may facilitate the prevention of multiple organ dysfunction syndromes.
Insights
Somatostatin (SST) treatment in macaques reduced Peyer's patches (PP) size during multiple organ dysfunction syndrome (MODS). SST also boosted B-cell immunity, potentially preventing MODS by shifting immunity from natural to acquired forms.
Area of Science:
- Immunology
- Gastroenterology
- Endocrinology
Background:
- Peyer's patches (PP) are crucial for intestinal immunity.
- Multiple organ dysfunction syndrome (MODS) significantly impacts immune function.
- The role of somatostatin (SST) in modulating immune responses within PP during MODS is not fully understood.
Purpose of the Study:
- To investigate the effects of somatostatin (SST) on immune B cells within macaque intestinal Peyer's patches (PP) during MODS.
- To elucidate the underlying mechanisms of SST's action on these immune cells.
Main Methods:
- 15 healthy adult macaques were divided into control, MODS, and MODS+SST groups.
- MODS was induced via superior mesenteric artery (SMA) occlusion and reperfusion (IIR).
- SST was administered prophylactically in the MODS+SST group; immune cell markers (CD20, CD5, TLR4, TLR2, α4β7, MadCAM-1) and PP morphology were assessed via immunohistochemistry.
Main Results:
- MODS increased the number and size of PP; SST treatment reduced PP number but not morphology.
- SST administration significantly increased CD20+ B-cells and plasma cells compared to MODS.
- SST downregulated TLR4 and TLR2 expression while upregulating MadCAM-1 in PP during MODS.
Conclusions:
- Endogenous somatostatin (SST) regulates the balance between natural and acquired immunity in PP B-cells.
- Prophylactic SST administration shifts excessive natural immunity towards beneficial acquired humoral immunity.
- SST shows potential in preventing multiple organ dysfunction syndromes (MODS) by modulating immune responses.
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