Colonic epithelial cells are a major site of macrophage inflammatory protein 3alpha (MIP-3alpha) production in normal colon and inflammatory bowel disease.

BACKGROUND AND AIM: Macrophage inflammatory protein 3alpha (MIP-3alpha) is a recently described lymphocyte directed C-C chemokine expressed predominately at extralymphoid sites, including the intestine. The aim of this study was to determine whether colonic epithelial cells produce MIP-3alpha and whether its expression is upregulated in inflammatory bowel disease. METHODS AND RESULTS: We found that interleukin 1beta and tumour necrosis factor alpha dose dependently stimulated MIP-3alpha production in Caco-2 and HT-29 intestinal epithelial cells. In cytokine treated Caco-2 and HT-29 cells, a significant increase in MIP-3alpha protein production was observed after three hours and continued for at least 24 hours. Analysis of colonic tissues by quantitative real time polymerase chain reaction and ELISA revealed significantly elevated MIP-3alpha mRNA levels (7.9-fold; p<0.05) and protein levels (8.9-fold; p<0.05) in Crohn's disease compared with controls or ulcerative colitis. MIP-3alpha immunoreactivity in normal colon and inflammatory bowel disease was principally associated with crypt and surface epithelial cells. Moreover, MIP-3alpha protein levels were elevated in primary epithelial cells isolated from patients with inflammatory bowel disease. CONCLUSIONS: These findings indicate that increased enterocyte MIP-3alpha production may play an important role in lymphocyte activation and recruitment to the colonic epithelium in Crohn's disease and ulcerative colitis.

Antigen processing and presentation by intestinal epithelial cells - polarity and complexity.

The mechanisms by which gut-associated lymphoid tissue (GALT) maintains a balance between oral tolerance and active immune response in the face of exposure to high antigen concentrations remains a central question in mucosal immunity. Here, Robert Hershberg and colleagues discuss the evidence that human intestinal epithelial cells function as antigen-presenting cells (APCs) capable of regulating T-cell responses in the intestinal mucosa

Correlation of in vitro T-cell and B-cell function with responses to childhood vaccines in children with human immunodeficiency virus infection.

We analyzed T-cell responses to mitogens and antigens and B-cell differentiation in response to T-cell dependent (TCD) and independent stimuli in 22 human immunodeficiency virus (HIV)-infected children (1 to 9 years of age) according to the presence of protective humoral immunity at a mean time of 18 months after vaccination with Haemophilus influenzae type b, hepatitis B, diphtheria, and tetanus vaccines. The 17 vaccine responders had a mean of 3.2 responses. However, their antibody levels were lower compared with healthy children. The 5 nonresponders had a mean of 0.84 responses. There were no significant differences between responders and nonresponders regarding age, Centers for Disease Control and Prevention (CDC) disease class, CDC immunologic class, serum immunoglobulin (Ig) levels, or in the use of antiretroviral therapy. However, responders tended to have higher age-adjusted absolute CD4 cell counts than nonresponders (p = 0.07). Nonetheless, there was no correlation between antibody levels and age-adjusted CD4 counts for each of the 4 TCD vaccines. Responders had conserved lymphoproliferative responses to mitogens and to candida antigen; 7 (41%) had normal responses to tetanus antigen. While nonresponders had some conserved responses to mitogens, only 1 had a response to antigen. Thirteen responders (77%) and only 1 nonresponder (20%) had normal responses to at least 2 of the 3 mitogens and 1 of the 2 antigens (p = 0.04). Although defects in B-cell differentiation were detected in both groups, they were profound and generalized in the nonresponders. Fourteen responders (82%) had at least 1 normal B-cell response compared with none of the 5 nonresponders (p = 0.002). There were no correlations between normal lymphoproliferative responses and age, CD4 counts, serum immunoglobulin G (IgG) levels, or the use of antiretroviral therapy. Immunologic function is important in the evaluation of HIV-infected children.

T cell antigen receptor V gene usage. Increases in V beta 8+ T cells in Crohn's disease.

Crohn's disease represents part of a spectrum of inflammatory bowel diseases characterized by immune regulatory defects and genetic predisposition. T cell antigen receptor V gene usage by T lymphocytes was investigated using four MAbs specific for various V gene products. One MAb (Ti3a), reactive with V beta 8 gene products, detected increased numbers of T cells in a subset of Crohn's disease patients as compared with normal controls and ulcerative colitis patients. In family studies there was no apparent inherited predisposition to the use of V beta 8 genes, and there was no association between a restriction fragment length polymorphism of the V beta 8.1 gene and Crohn's disease. The V beta 8+ T cells were concentrated in the mesenteric lymph nodes draining the inflammatory lesions and belonged to both the CD4+ and CD8+ T cell subsets. In contrast, lamina propria and intraepithelial T cells were not enriched in V beta 8+ T cells, suggesting that these cells were participating in the afferent limb of a gut-associated immune response. The expanded V beta 8+ T cells in Crohn's disease appear to result from an immune response to an as yet unknown antigen.

T cell antiidiotypic antibodies reveal differences between two human leukemias.

Two different human T cell leukemias were compared, using antiidiotype-like murine monoclonal antibodies. In each case these antibodies immunoprecipitated disulfide-linked heterodimer molecules from their respective leukemic cells. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the two idiotype-bearing molecules a major difference in molecular weight was observed, which could be attributed to a similar difference in size of the heavily iodinated chain of either heterodimer. The lightly iodinated chains of both molecules co-migrated at 43 Kd, but appeared to have different isoelectric points on two-dimensional gel analysis. The possibility that these two different heterodimers correspond to different classes of the putative T cell receptor for antigen is discussed. Assays of proliferation of the leukemic cells using Sepharose-bound antiidiotype-like monoclonal antibody showed that one of the leukemic cell types proliferated readily in response to its antiidiotypic antibody. This proliferation was not associated with measurable production of IL-2 and appeared to be a direct effect of the antiidiotypic antibody, which may mimic antigen in its interaction with the T cell receptor for antigen. The other leukemic cell type did not respond to Sepharose-bound antiidiotypic antibody and was generally unresponsive to lymphokines and mitogens. It is possible that the two leukemic cell types represent different stages of T cell differentiation.

Percutaneous choledochoscopy and cholecystoscopy: diagnostic and therapeutic uses.

Accessibility to the common bile duct and gallbladder through sinus tracts formed after placement of a T-tube or cholecystostomy tube at surgery has until recently been restricted to special catheters and accessories for the extraction of stones. The procedure requires continuous fluoroscopy while contrast media is injected in order to identify defects and to place accessories. More recently this techniques has been accomplished with endoscopes which are advanced into the bile duct through these tracts. The results are comparable to the catheter technique but exposure to x-ray is reduced. The procedure has been performed with a bronchoscope (BF-4B2-Olympus Corporation of America), modified with an irrigation-suction valve, permitting direct examination, biopsy and entrapment of stones. With the bronchoscope in the bile duct, physiologic function and pathologic conditions can be assessed by advancing catheters through the ampulla into the duodenum, 1) the former to facilitate manometric recordings and, 2) the latter to calibrate the opening of the papilla. A catheter passed antegrade through the bronchoscope into the duodenum can be used as a guide for the simultaneous performance of sphincterotomy while the duodenoscope is in position. These new applications have placed choledochoscopy and cholecystoscopy in the armamentarium for the evaluation and treatment of biliary tract disease.