Intraepithelial T-cell Research Articles

Mucosal immunity

Advances have been made in understanding specialized aspects of acquired mucosal immunity and the importance of cross-talk between host innate and acquired immune responses in host mucosal defense. These advances include elucidation of a molecular basis for the coordinated trafficking and retention of lymphocytes in intestinal sites, new insights into the possible extrathymic origin of mucosal T cells in the intestine, and the description of ligands that may be the targets of intraepithelial T-cell recognition. The past year has witnessed the characterization of an array of regulated epithelial cell mediators and responses important for host mucosal defense against enteric microbial pathogens and firmly establishes epithelial cells as an integral component of the mucosal immune network. Animal models continued to further understanding of intestinal inflammatory disease, and studies on tissue transglutaminase have generated significant new insights into the immunopathogenesis of human celiac disease.

Lymphocytes and dendritic cells in the normal uterine cervix: An immunohistochemical study

Objective: Assessment of the appearance, distribution and numerical density of immune cell populations in the normal human uterine cervix. Study design: Setting: University Hospital Gasthuisberg. Subjects: 29 healthy women undergoing total hysterectomy for non-cervical benign uterine disease. Analysis: Immunohistochemistry and morphometrical analysis on histological sections containing ectocervix, transformation zone and endocervix, using antibodies against the following antigens: HLA-DR, CD4, CD22, CD1a and CD8. Statistical analysis: Wilcoxon rank sum test. Results: Lymphocytes in the epithelial and stromal compartments are predominantly T-lymphocytes. Intraepithelial T-lymphocyte and Langerhans' cell densities and their distribution are not influenced by the menstrual cycle and are the same in both ectocervix and transformation zone. Conclusion: The wide variation of T lymphocyte subpopulations and Langerhans' cell densities in the normal epithelium of the uterine cervix is stressed. We are the first to present a large and well-defined control series, which is indispensable to study the effect of smoking and other factors on the cervical immune system.

Hierarchical Interactions of Control Elements Determine CD8α Gene Expression in Subsets of Thymocytes and Peripheral T Cells

CD4 and CD8 are crucial for the development and function of T cells. An intergenic deoxyribonuclease I hypersensitive site region (cluster CIII) directs expression in mature CD8 T cells only. Here, we show that two further independent regions from the CD8 gene locus in conjunction with cluster CIII restore transgene expression in appropriate immature thymocytes. Deletion of two of the intergenic cluster CIII DNaseI-HSS in homozygous mutant mice affects expression of CD8αα homodimers on intraepithelial T cells (IEL), particularly on the γδTCR + subset. Surprisingly, none of the thymocyte or peripheral αβTCR T cell subsets are affected by this mutation, indicating hierarchical activation of these elements within the different T cell subsets.

Oral mucosal changes in coeliac patients on a gluten-free diet.

Oral mucosal lesions or dental enamel defects may be the only presenting features of coeliac disease. A series of 128 patients with coeliac disease (CD) on a gluten-free diet (GFD), 8 patients with a newly diagnosed CD, and 30 healthy controls participated in a clinical and histopathological study of their oral mucosa. Oral mucosal lesions occurred in 71/128 GFD-treated CD patients. in 4/8 untreated and in 10/30 controls, and oral symptoms in 85/128, in 6/8 and in 10/30, respectively. Five CD patients had aphthous ulcers. Moderate to severe lymphocytic inflammation occurred in 36/117 and in 14/117 of the biopsy specimens of GFD-treated CD patients, in 1/8 and 2/8 of untreated CD patients, and in 3/30 and in 1/30 of controls, respectively. Intraepithelial T-cells were significantly more frequent in GFD-treated CD patients than in controls. There was no difference between untreated CD patients and controls. In the lamina propria of the GFD-treated CD patients, T-cells were more frequent than in the other groups. Mast cells were significantly more frequent in patients with GFD-treated CD. Nine GFD-treated CD patients had raised serum endomysium IgA antibody titres, although five of them reported to follow a strict GFD. A lack of strict compliance with a GFD may be related to the high prevalence of oral changes and symptoms. In addition, T-cell infiltration in the oral mucosa tends to increase with a longer duration of CD, independent of GFD-treatment. Clinically, it is important to study the oral cavity of patients suspected of having CD where the only clue to the disease may reside, since no less than 66% of the patients in this study had oral symptoms.

HIV-associated lymphoepithelial cysts and lesions: morphological and immunohistochemical study of the lymphoid cells.

To determine the morphology, immunophenotype and bcl-2 protein status of intraepithelial lymphocytes in HIV-positive lymphoepithelial lesions. Seventeen cases (from adults and children) of HIV-associated parotid and lung lymphoid lesions were examined. In addition, three lymphoepithelial cysts from HIV-negative patients were studied in parallel. Immunohistochemistry was performed on paraffin embedded tissue with the following antibodies: CD20, CD79a, CD3, CD4, CD8, bcl-2, CAM5.2, AE1/3, MIB1, kappa/lambda light chains and EBV-LMP-1. Heavy chain rearrangement was sought by polymerase chain reaction (PCR) in four of the cases. The lymphocytes participating in lymphoepithelial lesions of HIV-positive patients had the morphology of centrocyte-like cells with occasional cells resembling centroblasts. The majority of these cells were of B-cell lineage, but occasional intraepithelial T-cells (CD8 positive, CD4 negative) were also present. T-cells also formed a significant component of the infiltrative lymphoid cells outside the lymphoepithelial lesions. These were mainly CD8 positive, but very occasional CD4-positive T-cells were also noted. None of the cases showed light chain restriction and the four cases did not demonstrate heavy chain rearrangement by molecular biology. The interesting finding was the absence of bcl-2 expression by the intraepithelial lymphocytes. In contrast, the intraepithelial lymphocytes seen in the non-HIV setting were strongly bcl-2 positive. The majority of these were B-cells, and very occasional CD8 and CD4 positive T-cells formed the intraepithelial population. It is postulated that this finding is due to the HIV causing down-regulation of bcl-2 protein.

Blockage of T-cell costimulation inhibits T-cell action in celiac disease

Background & Aims: Celiac disease is an exemplary model of T cell–mediated pathology. Therefore, therapeutic approaches that target T cells may successfully control this disease. CTLA-4 immunoglobulin (CTLA-4Ig) can inhibit T-cell activation by blocking the engagement of CD28. We took advantage of this tool to define the pathogenic role of gliadin-specific T cells in the induction of celiac disease. Methods: Duodenal biopsy specimens from 7 treated celiac patients were challenged in vitro with gliadin and CTLA-4Ig or CD40-Ig. After 24 hours, the biopsy specimens were analyzed for the presence of characteristic modifications induced by gliadin challenge. Results: CTLA-4Ig down-regulated the expression of CD25, intercellular adhesion molecule 1, interleukin 2, and interferon gamma (stained lamina propria mononuclear cells/mm 2; P < 0.05) induced by gliadin challenge, caused apoptosis of gliadin-specific T cells (apoptotic T cells/mm 2; P < 0.05), and inhibited the production of antiendomysial antibody ( P < 0.01). However, it did not control intraepithelial T-cell migration ( P = NS) and Fas expression by enterocytes. Conversely, CD40-Ig only controlled production of antiendomysial antibody. Conclusions: In an organ culture model, CTLA-4Ig controls many but not all of the immunologic features of celiac disease. GASTROENTEROLOGY 1998;115:564-572

Lymphocytic gastritis in a child with hemophagocytic syndrome

Lymphocytic gastritis (LG) is an unusual form of chronic gastritis first described a decade ago in adult patients. The characteristic feature of LG is the surface intraepithelial T-cell lymphocytic infiltration, with relatively few lymphocytes in the lamina propria. Endoscopically, LG often correlates with the features of so-called “varioliform gastritis.” LG has seldom been reported in children, and then usually in association with celiac disease. We describe a case of LG in a 2-year-old child who died of a hepatitis A virus-induced hemophagocytic syndrome (HPS). LG was diagnosed retrospectively at the postmortem examination. The stomach showed diffuse thickening of mucosal folds with small erosions. On microscopic examination prominent intraepithelial infiltration of the gastric mucosa by T-cells was found, but the small bowel showed no villous atrophy, excluding the diagnosis of celiac disease. To the best of our knowledge, this is the first description of a case of LG associated with an infection-related HPS.

Activation features of intraepithelial γδ T-cells of the murine vagina

The epithelium of the murine vagina contains a resident population of γδ T-cells that expresses a homogenous V-4.Vδ1 TCR lacking N-region junctional diversity, implying that these T-cells recognize a very limited array of antigenic structures. The vaginal γδ T-cells express a pattern of surface markers characteristic of memory/effector T-cells that have previously been activated. Although vaginal γδ T-cells do not express the major costimulatory molecules CD28 and CD2, they do proliferate in response to a systemically delivered anti γδ TCR stimulus. Vaginal γδ T-cells contain mRNA that encodes the keratinocyte growth factor raising the possibility that these cells play a role in the repair of vaginal epithelium following injury. While the antigen recognized by the vaginal γδ TCR is unknown, a model is proposed which attempts to relate some of the unusual phenotypic features of vaginal γδ T-cells to the physiological injury and shedding of vaginal epithelium that occurs during the estrous cycle.

Intestinal T-cell lymphoma: a reassessment of cytomorphological and phenotypic features in relation to patterns of small bowel remodelling

Intestinal T-cell lymphoma (ITCL) is an uncommon entity among primary gastrointestinal lymphomas. In this study we evaluated tumours from 20 patients presenting with (n = 8) or without (n = 12) a history of coeliac disease (CD). Neoplastic lesions were composed of predominantly small (n = 4), small-to-medium (n = 2), medium/mixed-to-large (n = 7) or large and anaplastic (n = 7) cells. Different patterns of tumour growth and remodelling of the small bowel wall were observed. Pattern a (n = 4) was characterized by an intramucosal spread of small tumour cells with a small growth fraction. This pattern resembles mucosal inflammation in CD. In pattern b (n = 2), ulcerated solitary or multiple tumours composed of small to medium-sized cells were observed. The adjacent or distant mucosa showed a nearly normal architecture. In pattern c (n = 7), ulcerated lesions were composed of medium-sized to large cells. Mucosal flattening occurred in all segments infiltrated by lymphoma. In pattern d (n = 7), bowel remodelling was observed along the small intestine even at sites not affected by lymphoma. The main neoplastic lesions were composed of pleomorphic large or anaplastic cells frequently expressing the CD30 molecule. Intramucosal spread of a small epitheliotropic T-cell population was observed in the vicinity or even at distant segments of the small bowel. The demonstration of clonal rearrangements of T-cell receptor genes helped to trace widespread occurrence of this small intraepithelial neoplastic component. We suggest that different features of tumour cells such as the expression of activation antigens may contribute to the remodelling of small bowel mucosa. The addition of immunophenotyping data to macroscopic and microscopic features of specimens provided evidence that this uncommon lymphoma exhibits a spectrum in cytological composition and growth patterns. However, despite the considerable heterogeneity of the cases analysed, most of them shared a characteristic immunohistochemical profile (CD3+, CD8+/-, CD103+), further substantiating the view that ITCL is the neoplastic equivalent of an intraepithelial T-cell subset of the small intestine. This phenotype and the intraepithelial accumulation of lymphoma cells observed in the surviving mucosa are clues to the diagnosis of this clinicopathological lymphoma entity characterized by a broad range of morphological expressions.

The role of the thymus in intestinal intraepithelial T-cell development.

The role of the thymus in intestinal intraepithelial T-cell development.

4 T-cell responses and cellular immunity in coeliac disease

Increasing evidence points to a direct role for T cells in the mediation of the coeliac intestinal lesion. There is good evidence for increased local T-cell reactivity, manifest as increased in T-cell activation in the lamina propria and T-cell proliferation in the epithelial compartment. A likely scenario is that gluten elicits antigen-specific responses by lamina propria T helper cells, probably of the Th1 (inflammatory-mediator) subtype, leading to secretion of pro-inflammatory cytokines. Such cytokines may have direct effects on intestinal enterocytes, as well as mediating indirect effects by upregulation of MHC antigens and by enhancing the activity of cytolytic T cells. Although gluten-specific IEL responses have not been demonstrated by intraepithelial T lymphocytes (IELs), increasing evidence suggests that IELs can act as cytolytic effector cells and hence are likely to exert enteropathic effects under the influence of pro-inflammatory cytokines.

Extrathymic intestinal T-cell development: virtual reality?

Extrathymic T-cell development is a topic of considerable interest and debate, with important implications for the mechanisms of T-cell maturation and repertoire selection. Recent evidence has suggested that intraepithelial T lymphocytes (IELs) of the small intestine can mature and undergo selection in the absence of a thymus. However, IEL precursors are present in the thymus and IEL development is known to be influenced by the thymus. Here, Leo Lefrançois and Lyn Puddington discuss these data and suggest that the differentiation pathway of IEL precursors is dependent on whether or not a thymus is present.

Gastrointestinal lymphoma

Primary gastrointestinal lymphoma comprises a group of distinctive clinicopathological entities, most of which are not included in current lymph node-based lymphoma classifications. They may be of B- or T-cell type, with primary gastrointestinal Hodgkin's disease being extremely uncommon. Most low grade B-cell gastrointestinal lymphomas are of mucosa-associated lymphoid tissue (MALT) type, so called because they recapitulate the features of MALT rather than those of lymph nodes. Paradoxically, however, most MALT lymphomas arise in the stomach, which normally contains no organized lymphoid tissue. These gastric MALT lymphomas appear to arise in MALT acquired as a reaction to infection of the stomach by Helicobacter pylori and their growth can be influenced by eradication of this organism from the stomach. Low grade MALT lymphomas, which usually have a very favorable clinical course, may undergo high grade transformation; high grade tumours also may arise de novo and these probably also belong to the MALT group. Immunoproliferative small intestinal disease (IPSID) is a special form of MALT lymphoma with a restricted geographic distribution, which is characterized by synthesis of alpha heavy-chain immunoglobulin. Other gastrointestinal B-cell lymphomas include mantle cell lymphoma, which presents as lymphomatous polyposis, and Burkitt's or Burkitt-like lymphoma. Enteropathy (celiac disease)-associated T-cell lymphoma (EATL) is the most common primary gastrointestinal T-cell lymphoma. This is a clinically aggressive tumor that arises from the intraepithelial T-cell population, which is increased in celiac disease.

Unique properties of a cytotoxic CD4+CD8+ intraepithelial T-cell line established from the mouse intestinal epithelium.

Growth factor-dependent gut intraepithelial lymphocyte (IEL) cell lines were established from a long-term in vitro culture of BALB/c IEL with syngeneic irradiated spleen cells in the presence of concanavalin A-stimulated spleen supernatant fluids. The cell lines were preferentially consisted of very limited thymoindependent subsets of IEL; i.e., Thy-1+CD5-TCR alpha beta+CD4+CD8 alpha+beta- (double-positive; DP) IEL and Thy-1+CD5-TCR alpha beta+CD4-CD8 alpha+beta- (CD8 single-positive; CD8 SP) IEL. The CD8 SP IEL cell line had cytotoxic activities and was triggered to proliferate by T-cell receptor (TCR)-directed stimuli. The DP IEL cell line expressed high levels of the CD3-TCR alpha beta, exhibited cytotoxic activity in redirected lysis assays, and had perforin in the cytoplasm, indicating the functional maturity of this cell line. However, the DP IEL cell line did not proliferate in response to TCR alpha beta-directed stimuli, which indicated that TCR alpha beta-mediated signalling was able to initiate cytotoxic function but not to induce proliferation of the DP IEL cell line. Although both cell lines were shown to have functional competence, they expressed J11d antigen which marks immaturity in thymocyte differentiation pathways. These results indicate that the established thymoindependent DP and CD8 SP IEL cell lines have unique properties distinct from DP thymocytes and CD8 SP peripheral T cells. Together with a recent report on freshly isolated DP IEL (10), the unique properties of the DP IEL cell line seems to support the notion that DP IEL may undergo a unique maturation process in the gut microenvironment.

Molecular cloning of the human mucosal lymphocyte integrin alpha E subunit. Unusual structure and restricted RNA distribution.

The human mucosal lymphocyte-1 (HML-1) antigen is expressed on a subclass of T-lymphocytes known as intra-epithelial lymphocytes which are located between mucosal epithelial cells. The HML-1 complex is known to mediate adhesion of intra-epithelial T-lymphocytes to epithelial cell monolayers in vitro. We and others have shown that the HML-1 antigen is an integrin composed of the beta 7 subunit in association with a novel alpha subunit, alpha E. Here we report the cloning of the alpha E cDNA and its primary amino acid sequence. alpha E contained an inserted or I domain and was more homologous to the other I domain containing integrins than to the cleaved group of integrin alpha subunits. However, alpha E contained a unique extra domain of 55 amino acids located just NH2-terminal to the I domain without counterpart in other integrins. This extra domain contained a stretch of 18 consecutive charged residues and included a proteolytic cleavage site. Thus alpha E is the only I domain containing integrin alpha subunit that is also cleaved, and the cleavage site is distinct from that of members of the cleaved group of integrin alpha subunits. These structural features mark alpha E as an unusual member of the integrin family. High levels of alpha E and beta 7 mRNA were restricted to mucosal lymphocytes supporting the hypothesis that alpha E beta 7 plays a role in the localization or site-specific functions of intra-epithelial T-lymphocytes.

Analysis of Eimeria acervulina-induced changes in the intestinal T lymphocyte subpopulations in two chicken strains showing different levels of susceptibility to coccidiosis

The major T lymphocyte subpopulations expressing CD8, CD4, or antigen-specific T cell receptor ( TCR) heterodimer αβ (TCR2) or γδ ( TCR1) were assessed in duodenum intraepithelium of two inbred chicken strains, SC and TK, by single- and double-label immunofluorescence ( IF) at various times after oral inoculation with Eimeria acervulina. The duodenum intraepithelial T lymphocytes ( IEL) expressing the CD8 antigen (cytotoxic/suppressor T cells) increased in the SC and TK chickens following primary infection. However, a significant increase in the duodenum CD8 + IEL occurred in the SC chickens which manifested as a lower level of oocyst production compared to the TK chickens following primary and challenge infections. The CD4 + (T helper) cells increased seven days after primary infection in SC and TK chickens, and in TK chickens seven days after challenge infection. Two colour immunofluorescence analysis of duodenum IEL at 10 days after challenge infection revealed that the majority of CD8 + IEL expressed the TCR2 antigen. Furthermore, the SC chickens showed increased TCR2 +CD8 + cells shortly following challenge with E acervulina. These results suggest that variations in T cell subpopulations may reflect eimerian infection-related changes in the gut-associated lymphoid tissues and that a significant increase in CD8 + IEL in SC chickens may reflect enhanced acquired immune status in SC chickens compared to TK chickens.

Development of immune response markers in the trachea in the fetal period and the first year of life

Immunoglobulin (Ig)-producing cells, T cells (CD3) and epithelial expression of secretory component (SC) and HLA class II determinants (DR, -DP, -DQ) were studied by immunohistochemistry in 16 fetal and 15 postnatal specimens from the tracheal wall. Small amounts of secretory component (SC) was present in the tracheal surface and gland epithelium in the fetal period and increased towards term. A few IgM-, IgD- and IgG-producing cells were present in some fetal specimens but no IgA- and IgE-producing cells were found. Only very few CD3+ T-cells were present in fetal specimens and intraepithelial T-cells were virtually absent until after birth. Premature infants that lived for 1 week had less SC epithelial expression than mature infants of the same age. The density of CD3+ T-cells, IgA-, IgM-producing cells as well as the epithelial SC expression increased rapidly after birth. Epithelial MHC class II expression was absent in fetal specimens. HLA-DR was detected on the apical border of the surface epithelium one week after birth and was extensively expressed throughout the remaining postnatal period. Epithelial DP and PQ expression were virtually absent during this same period. These features probably reflect local activation of the immune system in response to environmental factors.

Intestinal intraepithelial lymphocytes and lymphoepithelial interactions in the human gastrointestinal mucosa.

Although representing a major immunological apparatus, it is not known how the immune system of the intestinal mucosa differentiates between dietary antigens (resulting in systemic tolerance) and potential pathogens. It is thought that intraepithelial T lymphocytes (IEL) may play a central role in local intestinal immunity and are likely to be important in immunity to gastrointestinal neoplasms and rejection responses to gut allografts. However, the biology of IEL and their unusual immunological microenvironments in the gastrointestinal mucosa are little understood. IEL are predominantly CD8+ TcR alpha beta+ CD3+ T cells which differ from lamina propria and peripheral T cells in many respects. IEL show low expression of CD5, CD6, LFA-1 (CD11a/CD18) and VLA-4, and high expression of HML-1. TcR gamma delta + IEL, although a minority population, are also phenotypically distinct, insofar as they are 50% CD8+, mainly V delta 1+ V gamma 9- and CD4- CD5-. IEL show poor proliferative responses to PHA, anti-CD3 and phorbol ester/calcium ionophore in vitro and have no clear functional role: they neither provide helper nor suppressor functions for Ig synthesis by B cells and do not mediate spontaneous cytotoxicity. However, there is evidence that IEL show preferential activation in response to sheep erythrocytes, presumably signalling via CD2. As normal and inflamed intestinal epithelia do not express ICAM-1, it seems unlikely that the LFA-1/ICAM-1 interaction is of importance to IEL activation. Rather, the CD2 (LFA-2) interaction with LFA-3 expressed by enterocytes may serve both to anchor IEL and to provide an accessory stimulus for activation. Nevertheless, the questions of antigenic specificity and immunological role remain unanswered.

Expression of the intestinal T‐lymphocyte associated molecule HML‐1: analysis of 75 non‐Hodgkin's lymphomas and description of the first HML‐1 positive T‐lymphoblastic tumour

The expression of the gut intra-epithelial T-cell associated molecule HML-1, a trimeric protein of 150, 125, 105 kD, was studied in 75 T-cell lymphomas of different subtypes: 20 T-lymphoblastic lymphomas/leukaemias; 50 nodal peripheral T-cell lymphomas; and five intestinal T-cell lymphomas. Our results confirm: (i) the usefulness of the HML-1 monoclonal antibody as an immunohistochemical marker for intestinal T-cell lymphomas: and (ii) the lack of reactivity of HML-1 with nodal peripheral T-cell lymphomas. Moreover, expression of the HML-1 molecule was found for the first time in a case of T-lymphoblastic lymphoma/leukaemia. The patient presented with a mediastinal mass which consisted of HML-1 + neoplastic cells displaying a phenotypic profile consistent with early thymocytes. Genes coding for the alpha, beta, gamma and delta chains of the T-cell receptor were in a germline configuration. The neoplastic cells could have been derived from the small subset of HML-1 + thymocytes detectable in the cortex of normal human thymus.

Cœliac disease 40 years gluten-free

History of coeliac disease. Dicke and the origin of the gluten-free diet.- HLA and coeliac disease, a finnish family study.- Adenovirus 12 and coeliac disease.- Gliadin peptides: their effect on the gut mucosa.- HLA-DR-DP and -DQ antigens in coeliac disease.- A cis- or transassociated DQ heterodimer predisposes to the gluten sensitive enteropathy seen in coeliac disease and dermatitis herpetiformis.- Nonhuman primates as animal models for coeliac disease: approach and feasibility.- T-cell subpopulations in intestinal mucosa.- Persistent increase of intra-epithelial T-cell receptor ? ? cells in coeliac disease.- T-cell subpopulations in coeliac disease longterm follow up with a gluten containing diet.- Intraepithelial T-cells of the TcR?/?+CD8- and V?1+ phenotypes are strikingly increased in coeliac disease.- Intestinalg amma/delta receptor bearing T lymphocytes are increased in coeliac disease and dermatitis herpetiformis.- Detecting gluten and related prolamins in food.- Gluten-free products, the Dutch experience.- The quest for gliadin limits and tolerance.- The incidence of coeliac disease and changes in gluten consumption.- Cath-up growth in 60 children with coeliac disease.- Clinical features of adult coeliac disease in Italy.- Associated diseases in children with coeliac disease.- Coeliac disease and autoimmune enteropathy.- Coeliac disease, malignancy and the effect of gluten free diet.- Dermatitis Herpetiformis: coeliac disease of the skin.- Latent coeliac disease.- Small intestinal biopsy.- High incidence of atypical forms of coeliac disease in a pediatric case study.- Coeliac disease in The Netherlands.- IGA reticulin antibody binding to human jejunum in sera of children with coeliac disease: the possible autoantibody of gluten sensitive enteropathy.- Evidence for a dominant gene mechanism in coeliac disease.- Crypt intraepithelial lymphocytes number is a sensitive index of dietary compliance in teenager coeliacs.- Upper G.I. damages seen at endoscopy in children with coeliac disease.- HLA class II contribution to coeliac disease -a family study-.- Diabetes-related immunologic markers in children coeliac disease.- HLA antigens and phenotypes associated with severe immunoglobulin - a deficiency in coeliac disease paediatric patients.- DR3 and DR7 -negative coeliac disease- a clinically relevant entity?.- Intestinal expression of HLA-DQ alleles associated to coeliac disease.- Is persistent adenovirus 12 (Ad12) infection involved in coeliac disease?.- Negative humoral response to adenovirus 12 in coeliac disease.- Characterization of ? ? TCR in T cell clones from small intestine of coeliac disease patients.- Characterization of intestinal ? ? and ?? T-cell receptor positive cells in coeliac disease.- Intercellular adhesion molecule-1 expression in coeliac disease.- Coeliac disease in Cyprus.- Coeliac disease in saudi children.- The spectrum of gluten intolerance in a defined paediatric population.- The reasons of malnutrition in children with coeliac disease.- Nervous system involvement in paediatric coeliac patients.- Coeliac disease and epilepsy with posterior cerebral calcifications: a causal association.- Genetic and immunological markers in children with dermatitis herpetiformis.- The clinical use of the gliadin antibody test.- Serum antigliadin antibodies during gluten challenge.- IgG-subclass antibodies to food antigens in patients with coeliac disease.- Isolation and characterization of wheat proteins with antibody reactivity against serum of a patient with coeliac disease.- Specific IgA-gliadin antibody as a marker of relapse in coeliac children on gluten challenge.- Gluten and dermatitis herpetiformis clinical observations in 5 active DH patients on an elemental diet.- Serum transglutaminase in the follow-up of coeliac disease.- Methods for measurement of gluten in foodstuffs.- Identification of coeliac-active peptides using a novel in vitro test for cellular immunity.- Unresponsive subtotal villous atrophy, ulcerative jejunitis: treatment with omeprazole.- Acknowledgements.