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T S Halstensen

Publications and source records attributed to T S Halstensen.

At least 37 records · Page 2Linked to original sources
PubMed1995

Human intestinal B-cell blasts and plasma cells express the mucosal homing receptor integrin alpha 4 beta 7.

Interactions between homing receptors on circulating leucocytes and endothelial addressins regulate tissue-specific cellular extravasation. Although integrin alpha 4 beta 7 appears to be the main receptor for gut-homing T lymphocytes, less is known about molecules mediating mucosal B cell homing. Expression of integrin alpha 4 beta 7 on B lymphocytes, B cell blasts, and plasma cells in human gut-associated lymphoid tissue (GALT; the Peyer's patches and appendix) and lamina propria was studied by multi-colour immunofluorescence applied on cryosections. Isolated mononuclear cells from the same tissue compartments were examined by flow cytometry and compared with peripheral blood B cells. Integrin alpha 4 beta 7 was expressed by IgA+ B cell blasts and plasma cells (CD38high) in the lamina propria, B cell blasts in GALT, and sIgD+ B lymphocytes in peripheral blood. In contrast, GALT sIgD+ B lymphocytes were negative or only weakly positive for alpha 4 beta 7. These results suggested that B lymphocytes down-regulate alpha 4 beta 7 upon extravasation in GALT but up-regulate this integrin after antigen-priming. Thus, alpha 4 beta 7 may be a homing receptor also for B cell blasts extravasating in the gut lamina propria, where this integrin is maintained on plasma cells, perhaps as a local retention factor.

ADP-ribosyl Cyclase↗
PubMed1995

Epithelial deposits of immunoglobulin G1 and activated complement co-localize with the "M(r) 40kD" putative auto-antigen in ulcerative colitis.

In conclusion, the "M(r) 40kD" antigen is expressed on the apical face of colonic enterocytes often in spatial relation to immune complexes in active UC, apparently targeting an IgG1-mediated autoimmune attack. Thus, our findings support the notion that an autoimmune response to the "M(r) 40kD" antigen, with local production of specific IgG1, is a possible immunopathological mechanism(s) in UC.

Autoantigens↗
PubMed1995

TCR gamma/delta + and CD8+TCR alpha/beta + intraepithelial lymphocytes (IEL) express proliferation marker (Ki-67) in the coeliac lesion.

In conclusion, it appears that gluten induces a non-proliferative activation of CD4+ lamina propria T cells, but a proliferative activation in the TCR alpha/beta +CD8+ and TCR gamma/delta + IEL subsets. Although the increased density of TCR alpha/beta +CD8+ and TCR gamma/delta + IEL in the coeliac lesion may be secondary to T cell mediated immune activation in the lamina propria, a direct and primary antigen-driven proliferation of IEL (perhaps CD1-dependent29) cannot be excluded.

Adolescent↗
PubMed1995

[Anders Jahre Prize for young researchers 1994. Chronic intestinal inflammation studies with multi-colored immunohistochemistry].

Multi-colour immunofluorescence can be a potent tool in investigating the cause of chronic inflammation of unknown etiology. The immune system uses two main mechanisms for target destruction; antibody-induced, complement-mediated or cell-mediated lysis. Immunohistochemical tracing of complement activation and immunoglobulin deposition in intestinal lesions from patients with ulcerative colitis and Crohn's disease show that ulcerative colitis may be an autoimmune disease characterized by an autoantibody of the IgG1 subclass to an apically located colonic autoantigen, which induce in situ complement activation and cellular lysis. The apical immune complex deposits co-localized with a previous identified putative autoantigen of 40kD, with peptide sequence homology with tropomyosin. Coeliac disease is a mucosal hypersensitivity to the wheat protein gluten. The intestinal lesion shows a peculiar disease specific increased percentage of T cell receptor (TCR)gamma/delta cells in the epithelium; differential use of CD45-isoforms and proliferative activation of intraepithelial T cells. T-helper(CD4) cells in the lamina propria show a non proliferative activation (CD25) which can be induced after gluten challenge in vitro. Immunomagnetic isolation of such in vitro activated T cells renders it possible to establish gluten reactive T cell clones that respond to gluten presented in the context of the disease associated human leucocyte antigen HLA-DQ2.

Autoantibodies↗
PubMed1994

Monoclonal antibody EG2 does not provide reliable immunohistochemical discrimination between resting and activated eosinophils.

The monoclonal antibody (mAb) EG2 has been considered to identify activated eosinophils and several immunohistochemical reports of EG2+ eosinophils in various allergic and other inflammatory disorders have suggested an important pathogenic role for such cells. This study showed that cellular EG2 reactivity, both in peripheral blood and mucosal tissue preparations, depends mainly on the method of sample preparation. Nearly 100% of blood eosinophils from normal individuals were strongly EG2+ when prepared by formalin fixation, whereas only a fraction reacted in the unfixed (64%) or acetone-fixed (60%) state. A significantly increased (p < 0.03) number of EG2+ cells were likewise detected in cryo-sections of inflamed nasal mucosa after formalin fixation compared with acetone fixation. Moreover, virtually all eosinophils were EG2+ in cryo-sections of normal jejunal mucosa fixed in periodate-lysine-(0.5%) paraformaldehyde prior to freezing. Conversely, EG2 reacted only weakly, or failed to react, with many eosinophils in cryo-material not subjected to such pre-fixation, in contrast to adjacent non-eosinophilic cells which were brightly stained. Two-colour immunofluorescence consistently revealed overlapping labelling with EG2 and mAb EG1 or a polyclonal antibody to eosinophil cationic protein in sections of formalin-fixed, paraffin-embedded normal gastrointestinal mucosa. Our findings thus showed that EG2 does not provide reliable immunohistochemical discrimination between resting and activated eosinophils. When optimal pre-fixation of tissue specimens was omitted, EG2 reactivity appeared to be caused, at least in part, by leached antigen adsorbed to adjacent non-eosinophilic cells.

Adult↗
PubMed1994

Differential interference contrast microscopy combined with immunofluorescence: a new method to phenotype eosinophils in situ.

Several surface receptors are expressed on eosinophils in vitro depending on the state of cellular activation, but immunohistochemical studies of eosinophil-related diseases have mostly focused on the number of infiltrating eosinophils as well as extracellular deposits of eosinophil granule proteins. The present investigation showed that eosinophils display a characteristic granular appearance in cryo-sections and cytospins by differential interference contrast (DIC) imaging. This approach appeared to be more reliable for identification of these cells in situ than immunohistochemical labelling of eosinophil granule proteins. Moreover, combined with immunofluorescence microscopy DIC imaging facilitated three-colour immunofluorescence phenotyping of eosinophils.

Cell Count↗
PubMed1994

Increased macrophage subset in inflammatory bowel disease: apparent recruitment from peripheral blood monocytes.

Mucosal specimens from active Crohn's disease (ileum, n = 6; colon, n = 6), active ulcerative colitis (n = 9), normal ileum (n = 6), and normal colon (n = 6) were subjected to paired immunofluorescence staining for characterisation of macrophage subsets in situ. In the normal state, only few CD68+ macrophages (< 10%) expressing the myelomonocytic L1 antigen (calprotectin) were seen. In inflamed mucosa, especially near small vessels, the CD68+L1+ fraction increased with the degree of inflammation, near ulcers to median 65% (range 35-91%). Cells reactive with the monoclonal antibody RFD7 were also increased in inflammation but less than 5% of them costained for L1 antigen. It is concluded that L1 producing macrophages are distinct from the RFD7+ subset and probably recently recruited from peripheral blood monocytes. Like granulocytes, L1+ macrophages may be important in non-specific defence, providing calprotectin with putative anti-microbial and anti-proliferative properties.

Adult↗
PubMed1994

Heterogeneity of M-cell-associated B and T cells in human Peyer's patches.

The specialized M cells in the follicle-associated epithelium (FAE) of Peyer's patches (PP) represent an intimate interphase between luminal antigens and gut-associated lymphoid tissue (GALT). M cells form pockets that contain clusters of leucocytes probably involved in the first encounter with antigens from the gut lumen. Three-colour immunofluorescence in situ phenotyping of these leucocytes in humans revealed about equal numbers of B (CD19/20+) and T(CD3+) lymphocytes, the latter mainly CD4+ (median 73%, range 40-90%), but relatively few macrophages (CD68+). Most B cells (90%) were positive for surface IgM (sIgM) and often co-expressed sIgD (median 34%, range 6-60%). Occasional B cells (median 2%) did not express CD45RA (range 0-15%) and 13% virtually lacked HLA-DR (range 0-40%). Some B and T lymphocytes expressed the nuclear proliferation marker Ki-67 (range 1-10%). The M-cell pockets also contained occasional cells with cytoplasmic IgA or IgM. These sites thus contained a heterogeneous B-cell population with features of both follicular mantle (sIgD+ sIgM+) and marginal zone (sIgD- sIgM+) B lymphocytes. Adjacent T lymphocytes were generally of the memory phenotype (CD45RO+). Our findings suggest that the M-cell-associated B lymphocytes represent local extensions of B-cell follicles towards the gut lumen, developed topically to facilitate antigen presentation and diversification of mucosal immune responses.

Adolescent↗
PubMed1993

Gliadin-specific, HLA-DQ(alpha 1*0501,beta 1*0201) restricted T cells isolated from the small intestinal mucosa of celiac disease patients.

Celiac disease (CD) is most probably an immunological disease, precipitated in susceptible individuals by ingestion of wheat gliadin and related proteins from other cereals. The disease shows a strong human HLA association predominantly to the cis or trans encoded HLA-DQ(alpha 1*0501,beta 1*0201) (DQ2) heterodimer. T cell recognition of gliadin presented by this DQ heterodimer may thus be of immunopathogenic importance in CD. We therefore challenged small intestinal biopsies from adult CD patients on a gluten-free diet in vitro with gluten (containing both gliadin and other wheat proteins), and isolated activated CD25+ T cells. Polyclonal T cell lines and a panel of T cell clones recognizing gluten were established. They recognized the gliadin moiety of gluten, but not proteins from other cereals. Inhibition studies with anti-HLA antibodies demonstrated predominant antigen presentation by HLA-DQ molecules. The main antigen-presenting molecule was established to be the CD-associated DQ(alpha 1*0501, beta 1*0201) heterodimer. The gluten-reactive T cell clones were CD4+, CD8-, and carried diverse combinations of T cell receptor (TCR) V alpha and V beta chains. The findings suggest preferential mucosal presentation of gluten-derived peptides by HLA-DQ(alpha 1*0501, beta 1*0201) in CD, which may explain the HLA association.

Amino Acid Sequence↗
PubMed1993

Activated T lymphocytes in the celiac lesion: non-proliferative activation (CD25) of CD4+ alpha/beta cells in the lamina propria but proliferation (Ki-67) of alpha/beta and gamma/delta cells in the epithelium.

In order to identify any dominating subset of activated T cells in the celiac lesion, we examined CD3+, CD4+, CD8+ and T cell receptor (TcR) gamma/delta+ lymphocytes in jejunal cryosections from 25 patients with celiac disease and 10 controls by three-color immunofluorescence staining for expression of the nuclear proliferation marker detected by monoclonal antibody (mAb) Ki-67 and the p55 alpha chain of interleukin-2 receptor (CD25). mAb Ki-67+ intraepithelial lymphocytes (IEL) were exclusively observed in celiac patients. The median proportion of CD3+ IEL positive for Ki-67 increased from nil in controls to 4.5% in partly treated (range 0-19.0%; n = 10; p = < 0.05) and 12.8% in untreated celiac disease (range 4.0-30.7%; n = 15; p < 0.005). Only 1.5% of CD3+ subepithelial T cells expressed the Ki-67 marker in celiac disease (range 0-9.5%). Two- and three-color staining combining mAb to CD3 and Ki-67 with mAb to CD4, CD8 or TcR delta showed that both TcR alpha/beta+ CD8+ and TcR gamma/delta+ (but not CD4+) mucosal T cells proliferated in the epithelium. By contrast, CD25 were almost exclusively expressed on CD4+ T cells in the lamina propria. The percentage of CD25+ T cells increased significantly from 1.7% in controls (range 0-2.9%) to 7.5% in partly treated (range 0.8-17.8%, p < 0.002), and to 14.65% in untreated celiac disease (range 3.9-21%, p < 0.002). These results suggest that gluten ingestion in celiac disease induces proliferative activation of TcR alpha/beta+ CD8+ and TcR gamma/delta+ IEL but non-proliferative activation (lymphokine production?) of lamina propria CD4+ T cells.

Adolescent↗
PubMed1993

Distribution and phenotypes of duodenal intraepithelial gamma/delta T cells in patients with various types of primary B-cell deficiency.

Expression of the gamma/delta T-cell receptor (TCR) on CD3+ intraepithelial lymphocytes (IEL) was studied in situ by two-color immunofluorescence on duodenal tissue sections from 34 infection-prone, adult patients with various types of primary hypogammaglobulinemia, classical Bruton's, Bruton-like or congenital, and common variable immunodeficiency. TCR gamma/delta+ IEL proportions (median 4.3%, range 0.3-43.3%) were within the range (0.3-38.3%) for histologically normal controls (n = 11), and there was no significant difference between the three patient categories. The total number of CD3+ IEL (mostly CD8+) per intestinal length unit was significantly higher (P < 0.004) in patients than in controls. In addition, TCR gamma/delta+ IEL per length unit, as well as TCR gamma/delta+ IEL proportions, were significantly increased (P < 0.008 and P < 0.05) in 14 patients with intestinal villous atrophy. Paired staining revealed that most (approximately 94%) TCR gamma/delta+ IEL in B-cell deficiency were CD8-, and a large fraction (approximately 67%) expressed the V delta 1/J delta 1-encoded epitope. No relationship was found between CD3+ or TCR gamma/delta+ IEL and the number of CD3+, CD4+, CD8+, and B lymphocytes or the CD4:CD8 ratio in peripheral blood. TCR gamma/delta+ IEL thus appeared to maintain a normal distribution in B-cell deficiency, except for being increased in patients with villous atrophy. Enhanced T-cell-mediated immunity, as possibly reflected by the numerical increase of CD3+ IEL, might compensate for a deficient mucosal B-cell system.

Adult↗
PubMed1993

Increased epithelial expression of HLA-DQ and HLA-DP molecules in salivary glands from patients with Sjögren's syndrome compared with obstructive sialadenitis.

Salivary gland specimens from 10 patients with primary Sjögren's syndrome (pSS) were examined by two-colour immunofluorescence with various combinations of monoclonal and polyclonal antibody reagents of the following specificities: human leucocyte antigen (HLA) class I and II (DR, DP and DQ), CD3, CD45 (leucocyte common antigen), various cytokeratins, and factor VIII-related antigen. Tissue specimens from 10 normal glands and 10 glands with obstructive sialadenitis (no known autoimmunity) served as controls. Only some intercalated ducts and scattered acini of the normal major glands expressed HLA class II determinants (< 5% of total epithelial area); the relative proportion of positive elements indicated differential expression (DR > DP > DQ). SS glands contained substantial T cell infiltrates and increased numbers of activated (DR+) T cells; adjacent epithelium showed extensive differential expression of HLA class II determinants (DR > DP > DQ). Glands with obstructive sialadenitis showed similarly increased epithelial expression of HLA-DR but with surprisingly small amounts of concomitant HLA-DP and -DQ expression. Epithelial HLA class II expression probably depends on cytokines as an inductive event, which is not unique for SS but particularly prominent in this disorder. Our results suggest that epithelial expression of HLA-DP or -DQ, rather than -DR, might be a prerequisite for the autoimmune process of SS to develop in genetically susceptible individuals.

Adult↗
PubMed1993

Duodenal intraepithelial gamma/delta T cells and soluble CD8, neopterin, and beta 2-microglobulin in serum of IgA-deficient subjects with or without IgG subclass deficiency.

Expression of the gamma/delta T cell receptor (TCR) on CD3+ intraepithelial lymphocytes (IELs) was studied by two-colour immunofluorescence in duodenal tissue sections from healthy (n = 6) or infection-prone (n = 7) subjects with selective IgA deficiency (IgAD), and subjects (n = 4) with combined IgAD and IgG subclass deficiency. TCR gamma/delta+ IEL proportions in selective IgAD subjects (median 6.3%, range 1.0-41%) and in those with combined deficiency (median 4.5%, range 1.2-33%) were well within the range (0.3-38%) for histologically normal controls (n = 11), but the healthy IgAD subgroup tended to show raised TCR gamma/delta+ IEL proportions (median 13.6%) compared with the other two subgroups. Also the number of TCR gamma/delta+ IELs per intestinal length unit was relatively high (median 13.9/mm) in the healthy IgAD subjects, and significantly raised (P < 0.03) compared with controls (median 3.2/mm). Paired staining revealed that most TCR gamma/delta+ IELs in both selective IgAD (98%) and combined deficiency (99%) were CD8-, and a large fraction (median 84% and 63%, respectively) expressed the V delta 1/J delta 1-encoded epitope. The total number of CD3+ IELs (mostly CD8+) was similar to controls. IgAD subjects, and especially the healthy subgroup, had significantly increased serum concentrations of soluble CD8 (P < 0.0002), neopterin (P < 0.005), and beta 2-microglobulin (P < 0.007), which was similar to our previous observations in common variable immunodeficiency, and probably reflected stimulation of cell-mediated immunity. In addition, the increased TCR gamma/delta+ IELs might reflect a component of compensatory surface protection in the healthy IgAD subgroup.

Adolescent↗
PubMed1993

Do human Peyer's patches contribute to the intestinal intraepithelial gamma/delta T-cell population?

T-cell receptor gamma/delta+) (TcR gamma/delta+) lymphocytes in human Peyer's patches (PP) adjoining ileal mucosa were studied by monoclonal antibodies with paired immunofluorescence staining in situ and by flow-cytometric phenotyping of isolated cells. The proportion of gamma/delta+ T cells in the follicle-associated epithelium outside the M-cell areas (median 4.1%, range 2.2-30.1%) was similar to that in mucosal villous epithelium (median 4.4%, range 0.5-30.5%). Most intraepithelial gamma/delta cells (approximately 90%) expressed CD45R0 but only a few expressed CD8 (< 10%) and none L-selectin; a dominating subset (median 46%) employed the V delta 1/J delta 1 gene product (range 22-100%). The M-cell areas lacked gamma/delta cells but contained clustered CD20+ and CD3+ lymphocytes. The subepithelial PP dome area and interfollicular (T-cell) zones, as well as the mucosal lamina propria, contained very few gamma/delta cells (median 1.7%, range 0.4-8.9%) which were dominated (88-100%) by the V delta 2-encoded subset. Those in the dome area and lamina propria were often (approximately 75%) CD45R0+ (range 44-90%) while very few (approximately 2%) expressed L-selectin (range 0-15%). By contrast, CD45R0 expression on gamma/delta cells in the PP T-cell zones was relatively low (approximately 46%) and that of L-selectin relatively high (approximately 43%). In conclusion, TcR gamma/delta + cells are quite rare in human PP and belong mainly to the V delta 2-encoded subset, thus being different from most intraepithelial gamma/delta cells that probably have another origin. The L-selectin+ fraction of PP gamma/delta cells presumably represent newly recruited 'naive' T lymphocytes while CD45R0+ gamma/delta cells both in PP and lamina propria are probably antigen-primed.

Adolescent↗
PubMed1993

Gluten stimulation of coeliac mucosa in vitro induces activation (CD25) of lamina propria CD4+ T cells and macrophages but no crypt-cell hyperplasia.

Jejunal biopsy specimens from 10 patients with treated coeliac disease and seven non-coeliac controls were challenged in vitro with peptic-tryptic gluten digest. Mucosal T cells were examined in situ by three-colour immunofluorescence staining for expression of the activation marker CD25 (the p55 alpha-chain of interleukin-2 receptor) and the nuclear proliferation marker revealed by monoclonal antibody Ki-67. Intraepithelial T cells expressed CD25 rarely whereas the proportion of activated lamina propria T cells increased (P < 0.002) from median 2.8% (cultured with 20% fetal calf serum alone for 24-48 h) to 10.0% after 24 h with gluten (n = 10; range 1.1-17.4%) and to 10.4% after 48 h (n = 7; range 1.4-17.5%). Such gluten-induced increase of CD25+ T cells was not observed in specimens from non-coeliac control subjects. Crypt-cell hyperplasia and T-cell proliferation (Ki-67+) were observed neither in the coeliac nor in the control mucosae after gluten stimulation. Three-colour staining combining a polyclonal antibody reagent to CD3 and a monoclonal antibody to CD25 with a monoclonal antibody to CD45R0, CD4, CD8, the p75 beta-chain of interleukin-2 receptor, integrin alpha E beta 7, or HLA-DR showed that most of the CD25+ T cells (> 90%) were CD4+CD8-, co-expressed CD45R0 and the p75 beta-chain, and often also the integrin alpha E beta 7 but not HLA-DR. In addition to these activated T cells, a dominating population of CD25+CD3-CD4+ subepithelial pan-HLA-class II+ macrophages (CD68+) with variable expression of the p75 beta-chain was often induced by gluten challenge.

Adult↗
PubMed1993

Distribution of macrophages and granulocytes expressing L1 protein (calprotectin) in human Peyer's patches compared with normal ileal lamina propria and mesenteric lymph nodes.

Antibodies to the cytosolic leucocyte L1 protein (or calprotectin) were examined for reactivity with macrophages, neutrophils, and eosinophils identified by paired immunofluorescence staining in sections of normal human ileal mucosa, including Peyer's patches. Macrophages were recognised by expression of the myelomonocytic antigen CD68 (monoclonal antibody KP1). Neutrophilic granulocytes were identified by their content of neutrophil elastase, and eosinophilic granulocytes by monoclonal antibody EG2. Virtually all CD68+ macrophages in normal lamina propria and Peyer's patches were L1- and the same was true for most extravasated macrophages in normal peripheral lymph nodes. Some mesenteric lymph nodes, however, and all peripheral lymph nodes with overt pathological processes (malignant lymphoma) contained many CD68+L1+ macrophages. Numerous L1+ cells were also localised to the crypt region and to some extent beneath the villous epithelium in normal lamina propria, but they were mainly identified as EG2+ eosinophils. Such cells were remarkably scarce or absent beneath the follicle associated epithelium in the dome region of Peyer's patches, where CD68+L1- macrophages were abundant. Also subepithelial and interfollicular CD68- interdigitating dendritic cells in Peyer's patches (recognised by antibody to S-100 protein) were usually unreactive with L1 antibody. The L1 protein shows a broad spectrum of antimicrobial activities in vitro, and its putative antiproliferative properties are interesting in relation to the immunosuppression postulated to take place in lamina propria. The virtual absence of L1 producing cells beneath the follicle associated epithelium in Peyer's patches may support the immunostimulatory function of these macrophage rich structures, which are held to be crucial for induction of specific mucosal immunity.

Adolescent↗
PubMed1993

Epithelial deposits of immunoglobulin G1 and activated complement colocalise with the M(r) 40 kD putative autoantigen in ulcerative colitis.

The intestinal expression pattern and general tissue distribution of the M(r) 40 kD putative epithelial autoantigen in ulcerative colitis were re-examined by in situ two and three colour immunofluorescence staining including the murine monoclonal antibody 7E12H12. The intestinal distribution was also compared with the epithelial codeposition of IgG1 and activated complement (C3b and terminal complement complex) seen selectively in ulcerative colitis. The M(r) 40 kD antigen was found for the first time in goblet cells of normal terminal ileum and proximal colon but not in rectal goblet cells. By contrast, colonic enterocytes expressed this antigen apically with increasing intensity in a distal direction, expanding to intense cytoplasmic expression in rectal enterocytes. The antigen was also expressed by the epithelium of the fallopian tubes, major bile ducts, gall bladder, and epidermis but not by proximal gastrointestinal tract epithelium or 13 other extra-gastrointestinal organs. Activated complement and IgG1 often colocalised with the M, 40 kD antigen apically on the surface epithelium in active ulcerative colitis but not in Crohn's disease. Our results support the idea that an autoimmune response to this antigen, leading to complement activation mediated by IgG1, is a possible pathogenetic mechanism for epithelial damage and persistent inflammation in ulcerative colitis.

Adolescent↗