Selective depletion of rectal lamina propria rather than lymphoid aggregate CD4 lymphocytes in HIV infection

The goal of this study was to examine the changes in lymphocyte populations in rectal mucosa during HIV infection and to study their relationship to mucosal immunity and to systemic depletion of CD4 lymphocytes. Rectal biopsies from 58 HIV-infected subjects and eight controls were studied. Frozen rectal tissue sections were stained with antibodies to CD4, CD3, CD8, and markers for macrophages. HIV-infected subjects were divided into early stage (no opportunistic infections) and AIDS groups. There was profound depletion of rectal lamina propria CD4 lymphocytes (16% and 6% of normal content in early and AIDS groups, respectively). However, lymphoid aggregate CD4 lymphocytes were far less severely depleted (69% and 40% of normal content, respectively). The extent of lymphoid aggregate CD4 lymphocyte depletion generally parallelled the CD4 lymphocyte depletion in the blood. CD8 lymphocyte content in both the lamina propria and lymphoid aggregates usually were increased, particularly in early-stage patients. Macrophage contents were usually normal in the HIV-infected groups. We conclude that rectal lamina propria and lymphoid aggregates are distinct compartments differing markedly in their CD4 lymphocyte content during HIV infection. In light of this and an increased number of apoptotic cells which were noted in rectal lamina propria in HIV-infected subjects, we hypothesize that intestinal lamina propria could be a site of rapid CD4 lymphocyte destruction during HIV infection.     

Mucosal lymphocyte subsets and HLA-DR antigen expression in paediatric Helicobacter pylori-associated gastritis

Paediatric studies may provide important insights into the immunopathology of Helicobacter pylori-associated gastritis, as mucosal changes reflect different stages of the immunoinflammatory response. We characterized, by quantitative immunohistochemistry, gastric mucosal lymphocyte phenotype and HLA-DR antigen expression and evaluated correlation with histopathology, in H. pylori-infected (Hp+ve) and uninfected children (Hp-ve). In the infected group, lamina propria CD3+ and IgA plasmocyte cell numbers were significantly higher and a trend for predominance of CD8+ over CD4+ was observed both in epithelium and lamina propria. A correlation of inflammation score with lamina propria CD3+ and CD4+ cell numbers and of CD45RO+ T lymphocytes with density of colonization was observed. The proportion of epithelial cells expressing HLA-DR antigen was significantly higher in the Hp+ve group and furthermore, glandular HLA-DR expression correlated with lamina propria CD3+ cell numbers, emphasizing the potential role of epithelial cells as antigen-presenting cells at this stage of infection.     

Functional analysis of TCR gamma delta+ T cells in tumour-infiltrating lymphocytes (TIL) of human pancreatic cancer.

The effect of HIV infection on intestinal lamina propria macrophage subsets was investigated in 41 patients at various stages of HIV infection (asymptomatic HIV infection, n = 17; AIDS, n = 24). Duodenal biopsies taken from HIV patients at endoscopy were snap frozen and cryostat sections cut for immunohistochemical staining. MoAbs CD68 (EBM11, pan-macrophage marker), RFD1 (antigen-presenting cells) and RFD7 (mature phagocytic macrophages) were used to identify cell subsets using indirect immunoperoxidase or alkaline phosphatase. Double immunofluorescence using MoAbs to HIV proteins (p24, p17 and gp120) and RFD1 were used to identify HIV-infected antigen-presenting cells. Double immunofluorescence was also used to identify macrophages that expressed both RFD1 and RFD7 ('suppressor' macrophages). Intensity of HLA-DR expression in lamina propria cells was investigated using a MoAb to HLA-DR directly conjugated to glucose oxidase. The results show that there was no difference in overall density of macrophages, but there was a significant decrease in dendritic cells (RFD1+) in all clinical stages of HIV. There was no difference in the density of RFD7+ macrophages, nor was there a difference intensity of HLA-DR expression in lamina propria cells. Only four HIV-infected cells were positively identified in the 41 patients. This result suggests that the antigen-presenting arm of mucosal immune defences may be seriously compromised in HIV infection, and represents a further insult to mucosal immunity already impaired as a result of loss of CD4+ T lymphocytes. This may contribute to development of opportunist infection in the gut.     

Analysis of intestinal lymphocyte subpopulations in patients with acquired immunodeficiency syndrome (AIDS) and AIDS-related complex

Lymphocyte subpopulations in the intestinal tissues of seven patients with acquired immunodeficiency syndrome (AIDS) or AIDS-related complex (ARC) were studied by immunohistologic technics at two different locations, the small bowel and the rectum. Intraepithelial and lamina propria lymphocyte subsets stained with monoclonal antibodies T11, T4, T8, and B1 were enumerated in the patients and different normal and patient control groups. Intraepithelial T11+ cells were decreased (P less than 0.05) in the small bowel of AIDS and ARC patients, primarily because of the near complete absence of T4+ lymphocytes. In the lamina propria of these patients, a depletion of T4+ cells (P less than 0.05), an increase in T8+ cells (P less than 0.05), and a reversal of the T4/T8 ratio were observed (e.g., the small bowel ratio was 0.1 +/- 0.02 vs. the normal ratio of 2.3 +/- 0.2 and the rectal ratios were 0.2 +/- 0.06 vs. normal 2.6 +/- 0.3). The T-lymphocytes in the intestine of AIDS and ARC patients did not express the receptor for interleukin-2 (IL-2). A near complete absence of T4+ lymphocytes was also seen in lymphoid follicles in the rectum. B1+ cells were not depleted. The reversal of the T4/T8 ratio, which is a hallmark of AIDS, occurs not only in the circulation but also in the gastrointestinal tissues of patients with AIDS and ARC.     

Infected cells and immune cells in the gastrointestinal tract of AIDS patients. An immunohistochemical study of 127 cases

Human immunodeficiency virus (HIV) proteins were detected by immunohistochemistry in the duodenal and rectal mucosa of 30% of 127 AIDS patients studied. HIV-infected cells were present in the lamina propria in 95% of the positive biopsies. They were immune cells, either isolated lymphocytes and macrophages (1-4 per positive biopsy) or dendritic reticulum cells forming a network in the germinal centres of mucosal lymphoid follicles. HIV proteins were not found in the duodenal epithelium or in the superficial rectal epithelium. In two cases (5% of the positive biopsies), they were found in rectal glands: the HIV-infected cells could be either epithelial cells or immune cells. This study confirms that the gut can be a target organ for HIV and that HIV is mainly carried by gut immune cells. The phenotypic study of lymphoid populations and macrophages in the gut mucosa of AIDS patients showed an inverse CD4/CD8 ratio in the lamina propria, compared with normal controls. This was independent of the presence of HIV proteins and is probably responsible for the appearance of opportunistic infections in the mucosa. An increase in activated macrophages was also noted in the mucosa of AIDS patients.     

The distribution of leucocyte subsets in the small intestine of healthy cats

The aim of this study was to investigate the distribution of leucocyte subsets in the small intestine of healthy adult cats (n=16). Immunohistochemical methods were used to identify leucocyte subsets within the mucosa of the duodenum, jejunum and ileum. Computer-aided morphometry was used to enumerate cells within the epithelial compartment, villous lamina propria and lamina propria adjacent to upper and lower crypt. Throughout the small intestine, IgA+ and IgM+ plasma cells were more prominent in the lamina propria adjacent to the lower crypt than in the villus, whereas IgG+ plasma cells were present in equal numbers in the crypt and villous regions. Overall, IgA+ plasma cells predominated and IgM+ plasma cells were higher in number than IgG+ plasma cells at each of the three anatomical locations. By contrast, T cells (CD3+) and T-cell subsets (CD4+ and CD8+) were present in greater numbers in the villous lamina propria than in the lamina adjacent to the crypts. Intraepithelial lymphocytes (IELs) were also characterized phenotypically, the majority being CD8+ T lymphocytes. Lamina propria macrophages and dendritic cells were characterized by expression of L1 and major histocompatibility complex (MHC) class II, and MHC class II expression by enterocytes overlying Peyer's patches, although rare, was also shown. The qualitative and quantitative data from this study provide a basis for comparison with cats with inflammatory enteropathies. Copyright Harcourt Publishers Ltd.     

Distribution of beta 7 integrins in human intestinal mucosa and organized gut-associated lymphoid tissue.

Two alternative integrins involved in mucosal homing (alpha 4 beta 7) or epithelial retention (alpha E beta 7) of lymphocytes were examined in the human gut. The distribution of the beta 7 subunit [monoclonal antibody (mAb) M301] was bimodal in that it was strongly expressed by alpha E beta 7 + cells but weakly by alpha 4 beta 7 + cells. More than 90% of intraepithelial lymphocytes (IEL), including the minor subsets of CD4+, T-cell receptor (TCR) gamma/delta +, and CD3- cells, expressed alpha E beta 7 as did most lamina propria CD8+ (88%) and a fraction (36%) of CD4+ lymphocytes. Conversely, B-lineage cells (CD19+) and macrophages (CD68+) were negative. In gut-associated lymphoid tissue (GALT: Peyer's patches and appendix) only a few (< 5%) cells were positive for alpha E beta 7 (confined to CD8+ lymphocytes and CD11c+ putative dendritic cells). A relatively small fraction of IEL (30-50%) expressed alpha 4 beta 7 (mAb Act-1), while most (70%) lamina propria T and B lymphocytes, blasts, plasma cells and macrophages were positive. In GALT, T lymphocytes expressed similar levels of alpha 4 beta 7 as in the lamina propria whereas relatively few B lymphocytes (< 50%) were positive. Isolated lamina propria CD8+, CD4+, CD19+, and CD38+ cells contained mRNA for alpha 4 and the former three subsets as well as appendix CD8+ cells also for beta 7 while only lamina propria CD8+ cells had mRNA for alpha E. Together, the results suggested that alpha E beta 7 and alpha 4 beta 7 are differentially regulated in inductive sites and effector sites of the human gut. Because lymphoid cells at both sites expressed mainly alpha 4 beta 7, this integrin may be a homing receptor on memory and effector cells bound for lamina propria as well as on naive lymphocytes extravasating in GALT. Conversely, because alpha E beta 7 was mainly expressed by CD8+ cells in epithelium and lamina propria, it was probably induced after extravasation, in agreement with the observation that IEL and a fraction of lamina propria T lymphocytes (mainly CD8+ cells) generally expressed higher levels of beta 7 than most CD4+ and B cells. Also a subset of putative dendritic cells located near the follicle-associated epithelium of GALT expressed alpha E beta 7, perhaps reflecting epithelial interaction during primary immune responses.     

Biological parameters of HIV-1 infection in primary intestinal lymphocytes and macrophages

Mucosal surfaces are the portal of entry for most HIV-1 infections and play an important role in disease pathogenesis. To characterize the biological parameters of HIV-1 infection in mucosal cells, we used purified lamina propria lymphocytes and macrophages from normal human small intestine to determine the distribution of the HIV-1 receptor and coreceptors on intestinal mononuclear cells and the permissiveness of these cells to HIV-1 infection. Lamina propria lymphocytes expressed CD4, CCR5, and CXCR4. In contrast, lamina propria macrophages expressed CD4 but not CCR5 or CXCR4. Intestinal lymphocytes supported replication by R5 and X4 isolates of HIV-1, but lamina propria macrophages were permissive to neither. RANTES, macrophage inflammatory protein-1alpha (MIP-1alpha), and MIP-1beta inhibited infection of intestinal lymphocytes by BaL, indicating that R5 infection of the intestinal lymphocytes was mediated by CCR5. Thus, resident lamina propria lymphocytes, not macrophages, are the target mononuclear cell for HIV-1 infection in the intestinal mucosa during early HIV-1 infection.     

Smoking influences the immunological architecture of the human larynx

Little is known about the effects of demographic and lifestyle factors on laryngeal mucosal immunology. Pinch biopsies of laryngeal mucosa were studied from 63 patients without laryngeal disease. Areas of positive staining for HLA-DR, HLA-DQ, HLA-DP, CD45, CD45RA, CD45RO, CD4, CD8, and CD79 were calculated. Patients were stratified according to gender and smoking status. Analysis of covariance showed current cigarette smokers had increased numbers of CD4+ T cells and there was an association between older age and greater CD4+ T cell numbers in both epithelium and lamina propria. Older age and female gender were associated with decreased lamina propria CD4+ CD45RO+ T cells and an increase in CD4+ CD45RO- T cells. T cell populations in the larynx may therefore be influenced by smoking, age and gender. We hypothesize that smoking induces changes in normal immunological function of the larynx, which may contribute to the etiology of inflammatory disease and cancer.     

Immunopathology of human immunodeficiency virus infection in the gastrointestinal tract

Conclusion The intestinal (in particular rectal) mucosa is an important portal of entry of HIV in homosexual men, who represent the vast majority of HIV-infected patients in Europe and North America. There are several possibilities for HIV to reach the CD4⁺ T cells, macrophages and follicular dendritic cells in the intestinal mucosa. HIV may be transported through M cells directly to mucosal lymphoid follicles. Alternatively, HIV may infect enterocytes via the Fc-receptor by antibody-bound HIV or via a CD4-independent receptor. By successive budding on the basal side of enterocytes HIV may be released into the lamina propria. Furthermore, in patients not infected by the intestinal route, HIV may also rapidly enter the intestinal mucosa by other mechanisms. Intestinal T lymphocytes are mainly activated memory T cells reentering the mucosal surfaces after circulating through the peripheral blood. In the periphery they may be preferentially infected by HIV. Accumulation of infected T cells could thus occur in the intestinal mucosa. The special phenotypical and functional characteristics of intestinal T lymphocytes may affect the replication and cytopathology of HIV, resulting in an accelerated loss of CD4⁺ T cells in the lamina propria. CD4 T cells play a critical role in antigen-dependent B cell differentiation, thus the pronounced CD4 T cell depletion in the intestinal mucosa may be responsible for the observed decrease of IgA plasma cells and a reduced secretion of IgA2. Depletion and functional impairment of activated mucosal LPL with consequent altered cytokine secretion in HIV infection could explain the breakdown of the mucosal immune barrier, leading to secondary opportunistic or nonopportunistic infections and secondary malignancies. Furthermore, several viral factors like thenef gene product may influence T cell activation and function, especially in the gastrointestinal tract contributing to the observed pronounced and early loss of CD4 T cells in the mucosal lamina propria. In addition, due to the interrelation between the mucosal immune system and epithelium, these changes might be responsible for the partial small intestinal mucosal atrophy and maturational defects in enterocytes observed in HIV-infected patients.     

Changes in natural immunity during the course of HIV-1 infection.

The role of natural killer (NK) and lymphokine-activated killer (LAK) cell-mediated cytotoxicity in AIDS has yet to be established. The objective of this study was to determine inducible LAK cell responses at different stages of HIV-1 infection, and specifically to establish the participation of CD8 lymphocytes in these responses. Peripheral blood lymphocytes (PBL) were isolated from healthy seronegative (CDC-0) subjects and HIV-1+ individuals who were clinically asymptomatic (Centre for Disease Control group 2, CDC-2) or symptomatic (CDC-4) with regard to secondary opportunistic infection (OI). LAK cells were generated upon incubation of PBL with IL-2 and their cytolysis of K562 and U-937 targets was determined using chromium release assays. The role of CD8+ lymphocytes as progenitors and effectors of these LAK cell responses was determined by immunomagnetic depletion of CD8+ cells from precursor PBL and LAK cells, respectively. LAK cell-mediated cytotoxicities in HIV-1-infected individuals were reduced compared with seronegative controls without any corresponding changes in the relative proportions of CD56+ (NK) cells among groups. Depletions of CD8+ subsets from either PBL or LAK cells dramatically reduced total LAK cytotoxic responses and LAK activities per unit CD56+ cell in the OI-/CDC-2 seropositive population. No corresponding changes in LAK activities in seronegative control or HIV+/OI+/CDC-4 groups were observed. Levels of LAK activity against K562 targets in CDC-0/HIV- and CDC-4/HIV+ groups correlated with the percentage of CD56+ LAK cells; corresponding LAK activity in the CDC-2/HIV+ group correlated with the percentage of both CD56+ and CD8+ subsets. These findings suggest that adaptive changes in non-MHC restricted cytotoxic responses occur in HIV-1 individuals at early stages post-HIV infection, before the onset of opportunistic infection.     

Strategies for optimizing targeting and delivery of mucosal HIV vaccines

Effective frontline defenses against HIV‐1 will require targeting vaccines to mucosal tissue in order to induce αβ CD8⁺ lymphocytes in mucosal effector sites (lamina propria and intraepithelial compartment) as well as antibody secreting plasma cells that can neutralize and limit free virus. A concerted second wave of assault against the virus will require the activation and recruitment of antigen specific memory CD4⁺ and CD8⁺ T cells in mesenteric lymph nodes and distal secondary lymphoid organs. New delivery strategies targeting the “right” DC subsets in combination with delivery of mucosal adjuvants and innate signals for activating DC will be essential for mucosal vaccines in order to circumvent the naturally tolerogenic environment and the induction of Tregs. Mucosal delivery of antigen in combination with inflammatory signals has been shown to empower systemic immunization by directing responses to mucosal sites for imprinting optimum mucosal memory. Here, we discuss novel vaccine strategies and adjuvants for optimizing mucosal delivery of HIV vaccines.     

T-cell subset alterations in HIV-infected homosexual men: NIAID Multicenter AIDS cohort study

Immunologic changes in HIV-infected homosexual men without AIDS were studied using flow cytometry and monoclonal antibodies. A decline in CD4 cells occurred after anti-HIV antibodies detectable by ELISA developed. CD4 T-cell levels dropped to an average of 60% of their original level within 12-18 months after seroconversion. Subsequently, CD4 levels remained constant in most HIV seropositive men for several years. However, in men who developed AIDS, there was a rapid fall in the CD4 level during the 2 years prior to development of AIDS. Throughout the course of HIV disease, the total T-cell levels (CD3) remained constant, apparently due to CD8 lymphocytosis. The selective depletion by HIV infection of discrete functional subsets of CD4 cells was examined using 4B4, 2H4, HB-11, and Leu-8 monoclonal antibodies and dual color immunofluorescence. No selective depletion of CD4 subsets was noted using any of these reagents. However, selective activation of subsets of CD8 lymphocytes characterized disease progression. In particular, increases in the number of HLA-DR+, CD38+ (OKT10), and Leu-8- CD8 lymphocytes were associated with a fall in CD4 levels and development of AIDS.     

Distribution of Alloreactivity Amongst the CD45 Isoforms of Circulating CD4 and CD8 T Lymphocytes

The expression of different isoforms of the CD45 surface molecule allows lymphocytes to be divided into two nonoverlapping categories, CD45RA and CD45RO. Previous studies of CD4 T cells have shown that responses to soluble antigens are present predominantly in the RO subset and to mitogens in the RA, alloreactivity being present in both subsets. Responses of CD8 T cells have not been investigated in such detail, nor have responses been compared to CD4 cells. Here we report the alloreactive responses of both CD45RA and RO subsets amongst both CD4 and CD8 T lymphocytes. Following isolation of CD4 and CD8 cells with immunomagnetic beads, CD45 subsets were separated by negative depletion using specific monoclonal antibodies. CD45RA populations were greater than 97% pure and CD45RO cells greater than 91%. One-way primary mixed lymphocyte reactions were established using the purified responder cells with irradiated allogeneic peripheral blood mononuclear cells as stimulators; experiments were all repeated at least three times. In assays of CD4⁺ RA and RO subsets, reactivity was present in both isoforms, being consistently, but not significantly, greater amongst the RO subset. With CD8⁺ T cells, reactivity was also present in both isoforms, but was significantly greater in the CD45RA subset, with mean proliferation 2.5–3-fold that of the CD45RO cells ( P  < 0.05).     

Immunobiology of the human penile urethra.

The human penile urethra contains numerous IgA and J chain-positive plasma cells, and the epithelium expresses secretory component, the transport molecule for polymeric IgA, indicating that this region is an active site of secretory IgA-mediated immune defense. At the distal tip, the mucosae of the meatus and fossa navicularis contain intraepithelial dendritic cells but few macrophages, whereas the urethra proper contains many macrophages within the lamina propria and epithelium, but no dendritic cells. T lymphocytes are abundant and ubiquitous in all regions of the urethra. Both CD8+ and CD4+ subpopulations of T lymphocytes are present in the lamina propria and epithelium, although CD8+ cells predominate. The majority of T lymphocytes are positive for CD45RO (memory marker), and many are also positive for the alpha E beta 7 integrin (mucosal-associated antigen). These data indicate that the human urethra is a highly dynamic immunocompetent tissue possessing all the necessary elements for antigen presentation and both humoral and cellular mucosal immune responses. Furthermore, the urethra resembles other mucosal surfaces in terms of lymphocyte subpopulations, segregation of phenotypes and expression of antigenic determinants characteristic of mucosal lymphocytes. It is likely that this region plays a dominant role in protecting the male urogenital tract against ascending infections, and should be targeted in vaccination strategies directed against sexually transmitted diseases.     

Human intestinal lamina propria lymphocytes are naturally permissive to HIV-1 infection

The presence of HIV-1 in the intestinal mucosa of AIDS patients has been reported and human intestinal lamina propria lymphocytes (LPL) have been proposed as important targets for HIV-1 infection. However, little information is available concerning the permissiveness of human intestinal CD4+ T lymphocytes to HIV-1 infection. Here, we show that human LPL, in contrast to autologous peripheral blood lymphocytes (PBL), are permissive to both X4 T-tropic and R5 M-tropic strains of HIV-1, as well as to clinical isolates, in the absence of exogenous stimuli. Flow cytometry showed that the vast majority of T LPL were CD45RO+ and CD69+, and that CD4+ T LPL highly expressed CC chemokine receptor 5 (CCR5) as compared to PBL, while CX chemokine receptor 4 was equally expressed on LPL and PBL. Exogenous RANTES and macrophage inflammatory protein-1alpha (natural CCR5 ligands) virtually abolished the entry of the R5 M-tropic strain HIV-1 into human LPL. Thus, we infer that human intestinal CD4+ T lymphocytes are naturally susceptible to HIV-1 infection, due to their physiological state of activation and to marked expression of HIV-1 coreceptors, independently of the route of primary (either mucosal or parental) infection and the shifts of the virus phenotype occurring during the course of AIDS.     

Human CD8+ intraepithelial T lymphocytes are mainly CD45RA-RB+ and show increased co-expression of CD45R0 in celiac disease

Expression of various CD45 isoforms (RA, RB and R0) on CD3+, CD4+ and CD8+ intraepithelial and lamina propria T cells was examined in situ by a three-color immunofluorescence technique in jejunal biopsy specimens from 32 patients with celiac disease and 18 controls. The median percentage of CD3+ intraepithelial lymphocytes (IEL) that expressed CD45R0 increased from 52% in controls to 69% in untreated celiac disease (p less than 0.01). Furthermore, the percentages of CD4+ and CD8+ IEL strongly positive for CD45R0 rose respectively from 94% and 24% in controls to 100% and 55% in untreated celiac disease. Conversely, CD45R0 was strongly expressed on most CD3+ lamina propria lymphocytes (LPL) both in control (81%) and diseased (77%-81%) mucosa. A variable fraction of the intraepithelial and lamina propria CD3+ T cells expressed mainly CD45RB (controls, 46% and 20%, respectively; celiac disease, 29% and 15%). Only 2% IEL and 4% LPL were positive for CD45RA. Expression of different CD45R isoforms thus identified three distinct CD8+ T cell subsets in human intestinal mucosa. In addition, our results suggested that antigen-primed CD8+CD45R0+ memory cells accumulate in the jejunal epithelium of patients with untreated celiac disease.     

Abnormalities in subset distribution, activation, and differentiation of T cells isolated from large intestine biopsies in HIV infection. The Berlin Diarrhoea/Wasting Syndrome Study Group.

Intestinal T cells have a unique state of activation and differentiation which might specifically affect or be affected by HIV infection. Lymphocyte subsets in the peripheral blood are well characterized, but our knowledge about intestinal lymphocytes in HIV infection is incomplete. We therefore analysed lymphocytes isolated from large intestine biopsies of AIDS patients and controls by three-colour cytofluorometry. In the large intestine of HIV-infected patients CD4 T cells were reduced and CD8 T cells were increased compared with controls. Most of the CD8 T cells in the colorectal mucosa of AIDS patients were of the cytotoxic phenotype. Activated and resting CD4 T cells were similarly reduced, the expression of CD25 and HLA-DR of CD8 T cells was unaltered and increased, respectively. In intestinal CD4 T cells the expression of CD29 was decreased, but the expression of CD45RO and HML-1 was normal. CD8 T cells had a decreased expression of all these differentiation markers. Our findings demonstrate substantial alterations in subset distribution, activation, and differentiation of large intestine T cells, which may contribute to the secondary infections and malignancies commonly observed in the gut of AIDS patients.     

Analysis of leucocyte subsets in the canine intestine

Immunohistochemistry and computer-aided morphometric analysis were used to define populations of leucocyte subsets in the intestinal tract of an outbred population of dogs with no evidence of gastrointestinal disease. In the small intestinal lamina propria, B cells and plasma cells (IgA+, IgM+ and IgG+) were prominent in peri-crypt regions, with a significant trend for a reduction in the number of cells towards the villous tip (P < 0.0001). By contrast, lamina propria T cells (CD3+) and T-cell subsets (CD4+ and CD8+) were present in greatest numbers at the tip of the villus, with significantly decreasing numbers towards the crypt regions (P < 0.0001). In the lamina propria, CD4+ cells outnumbered CD8+ cells (P = 0.05), but the opposite was true of the epithelial compartment (P < 0.001). The distribution of CD5+ lymphocytes was similar to that of CD3+ cells, in both the lamina propria and epithelial compartments. The numbers and distribution of cells expressing MHC class II, L1 and CD45 were recorded. Numerous eosinophils were present in the lamina propria, and an intra-epithelial population was also noted, especially in the crypt epithelium. Mast cells, which were mainly found in the subepithelial lamina propria, were also present within muscle layers, and cells expressing IgE had a similar distribution. Similar populations of cells were recorded in the colonic lamina propria and epithelium. The quantitative and qualitative data from this study will enable comparisons to be made with dogs suffering from inflammatory bowel diseases.