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Colonic spirochetosis caused by S. pilosicoli is a disease of human and animals characterized by intimate attachment of the spirochete to colonic epithelial cells and colitis. To identify antigens that are potentially involved in recovery from the disease, whole-cell lysate (WC) and various detergent extracts including Sarkosyl-soluble (SS) and insoluble (SI), and Triton X-114 detergent phase (TXD) and aqueous phase (TXA) of the human isolate SP16 were examined by Western blotting with Serpulina spp. periplasmic flagellar protein FlaB-specific monoclonal antibody 7G2 as well as pooled pre-immune serum (PS), hyperimmune serum (HS), and convalescent serum (CS) from swine. The HS reacted with several antigens that were not identified by the CS, including the periplasmic flagellar proteins and some lower molecular weight bands. The CS identified three major immunoreactive double (D) or single (S) bands of approximately: (i) 64-kDa in the WC(S), SS(D), and TXD/A(S), (ii) 54-kDa in the WC(S), SS/I(S), and TXD(S), and (iii) 47-kDa in the SS(S) fraction. The data indicate recovery from colonic infection elicits serum IgG antibodies to specific S. pilosicoli outer membrane antigens (SPOMA).

Transcription factors of the NF-kappaB/Rel family are critical for inducible expression of multiple genes involved in inflammatory responses. Sulfasalazine and its salicylate moiety 5-aminosalicylic acid are among the most effective agents for treating inflammatory bowel disease and rheumatoid arthritis. However, the mode of action of these drugs remains unclear. Here we provide evidence that the transcription factor NF-kappaB is a target of sulfasalazine-mediated immunosuppression. Treatment of SW620 colon cells with sulfasalazine inhibited TNFalpha-, LPS-, or phorbol ester- induced NF-kappaB activation. NF-kappaB-dependent transcription was inhibited by sulfasalazine at micro- to millimolar concentrations. In contrast, 5-aminosalicylic acid or sulfapyridine did not block NF-kappaB activation at all doses tested. TNFalpha-induced nuclear translocation of NF-kappaB was prevented by sulfasalazine through inhibition of IkappaBalpha degradation. When blocking proteasome-mediated degradation of IkappaBalpha, we could demonstrate that sulfasalazine interfered with IkappaBalpha phosphorylation, suggesting a direct effect on an IkappaBalpha kinase or on an upstream signal. Inhibition of NF-kappaB activation seems to be specific since other DNA-binding activities such as AP1 were not affected. These results demonstrate that sulfasalazine is a potent and specific inhibitor of NF-kappaB activation, and thus may explain some of the known biological properties of sulfasalazine.

The authors studied eight colectomy and eight biopsy specimens from 12 patients with cystic fibrosis who had developed fibrosing colonopathy, a complication observed in patients receiving high-strength enzyme replacement. The colectomies originated from five male and three female patients ranging in age from 18 months to 6 years. Five individuals had localized strictures of the right colon and three had stenosing fibrosis of the entire colon. The affected colon had a cobblestone appearance, submucosal fibrosis, thickening of the muscularis propria and chronic mucosal inflammation in all patients, with active cryptitis in four. Moderate to severe infiltration by eosinophils, with increase in the number of mast cells, and widespread interruption of the muscularis mucosa were present in every case. Four colectomies were preceded by endoscopic biopsies; four patients who have not undergone surgery also underwent biopsy. All the biopsies showed evidence of active or chronic inflammation, and all had increased mucosal eosinophils. Prolonged colonic mucosal contact with either the enzymes and/or the enteric coating itself may lead to mucosal colonic ulceration and inflammation. Topical allergy may then promote the stenosing fibroplasia.

Autoantibodies in chronic immune thrombocytopenic purpura occasionally interfere with platelet function. We describe a patient with a normal platelet count who had clinically significant mucosal bleeding, a prolonged bleeding time and abnormal platelet aggregation. The patient had high titres of an IgG4 kappa autoantibody, directed to a cation-dependent epitope on platelet glycoprotein IIb/IIIa, which blocked the binding of fibrinogen and fibronectin to this complex. Corticosteroid treatment resulted in clinical improvement and a marked drop in autoantibody concentration. The lack of thrombocytopenia in this patient, despite high autoantibody levels, is best explained by the poor recognition of IgG4 antibodies by phagocytic cells.

In a randomized study the effect of whole blood transfusion versus bedside leucocyte-depleted blood transfusion on lymphocyte proliferation, CD4+:CD8+ ratio, and levels of soluble interleukin 2 receptor (sIL-2R) and interleukin (IL) 6, as well as on the development of postoperative wound infection and intra-abdominal abscess, was assessed in 60 patients undergoing elective colorectal surgery. Transfusion with whole blood induced a significant decrease in lymphocyte proliferation and CD4+ :CD8+ ratio (P < 0.01) as well as a significant increase in sIL-2R and IL-6 levels (P < 0.01). Furthermore, transfusion with whole blood was accompanied by a significant increase in postoperative infectious complications (P < 0.01). In patients transfused with leucocyte-depleted blood only slight and transient changes were observed, which were not significantly different from those observed in non-transfused patients.

Gnathostomiasis is primarily a disease of the skin characterized as creeping eruption or mobile erythema. However, larval Gnathostoma sometimes migrate into an unexpected site to elicit serious illness. Here we describe a case of colonic ileus caused by Gnathostoma doloresi. The patient was a 57-year-old man living in Miyazaki Prefecture, Japan, which is known as an area endemic for this parasite. One week after having eaten a few slices of the flesh of a snake (Agkistrodon halys), he developed severe abdominal pain. An abdominal radiograph revealed multiple gas-fluid levels with a distended bowel of an inverted U shape. A barium enema revealed a tumor in the ascending colon near the hepatic flexure that was surgically removed by simple colonic resection. An oblique section of a parasite surrounded by massive infiltration of eosinophils was found by postoperative histopathologic examination. The entire body of the advanced third-stage larva of G. doloresi was dissected from a specimen-embedded paraffin block.

Background: Some cutaneous T-cell lymphoproliferative diseases (LPD), such as lymphomatoid papulosis and pityriasis lichenoides et varioliformis acuta, are characterized by an indolent or waning and waxing clinical course. However, such T-cell LPD are rarely documented in other organs., Methods: A patient with T-cell LPD of the digestive tract characterized by repetitive episodes of self-healing ulcers in the oral and intestinal mucosa over the course of 17 years is reported. Biopsy specimens from oral and intestinal mucosa were studied by conventional pathology, immunocytochemistry, and Southern blot analysis of T-cell receptor (TCR)-beta and -gamma gene rearrangement., Results: Immunocytochemically, the infiltrating lymphocytes were lamina propria T cells with a dominant phenotype CD3+, CD4+/-, CD8-, and HML-1-. DNA study revealed the same rearranged configuration of TCR-beta and -gamma genes in specimens from both oral and colonic lesions., Conclusions: The present case may represent a novel T-cell lymphoproliferative disease (i.e., a digestive-tract mucosal counterpart of cutaneous dysplastic LPD).

Pathogenic bacteria that penetrate the intestinal epithelial barrier stimulate an inflammatory response in the adjacent intestinal mucosa. The present studies asked whether colon epithelial cells can provide signals that are important for the initiation and amplification of an acute mucosal inflammatory response. Infection of monolayers of human colon epithelial cell lines (T84, HT29, Caco-2) with invasive strains of bacteria (Salmonella dublin, Shigella dysenteriae, Yersinia enterocolitica, Listeria monocytogenes, enteroinvasive Escherichia coli) resulted in the coordinate expression and upregulation of a specific array of four proinflammatory cytokines, IL-8, monocyte chemotactic protein-1, GM-CSF, and TNF alpha, as assessed by mRNA levels and cytokine secretion. Expression of the same cytokines was upregulated after TNF alpha or IL-1 stimulation of these cells. In contrast, cytokine gene expression was not altered after infection of colon epithelial cells with noninvasive bacteria or the noninvasive protozoan parasite, G. lamblia. Notably, none of the cell lines expressed mRNA for IL-2, IL-4, IL-5, IL-6, IL-12p40, IFN-gamma, or significant levels of IL-1 or IL-10 in response to the identical stimuli. The coordinate expression of IL-8, MCP-1, GM-CSF and TNF alpha appears to be a general property of human colon epithelial cells since an identical array of cytokines, as well as IL-6, also was expressed by freshly isolated human colon epithelial cells. Since the cytokines expressed in response to bacterial invasion or other proinflammatory agonists have a well documented role in chemotaxis and activation of inflammatory cells, colon epithelial cells appear to be programmed to provide a set of signals for the activation of the mucosal inflammatory response in the earliest phases after microbial invasion.

The relative proportions of cells containing the various classes of immunoglobulins in the colonic mucosa were determined in eight patients with human intestinal spirochaetosis (HIS) and in eight controls. All specimens were taken from colonic resections performed because of adenocarcinoma. None of the cases with HIS showed an inflammatory reaction in the mucosa as judged subjectively by light microscopy. Cases with HIS had smaller proportions of IgD- and IgE-positive cells and a larger proportion of IgA cells as compared with the controls, whereas the proportions of IgG and IgM cells were similar in the two groups. Taking into account the large individual variations in the proportions of immunoglobulins in both groups, our findings must be interpreted with caution. However, our results do not support the previous demonstration of increased percentages of IgE-positive cells in patients with HIS.

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.

Mucosal macrophages and accessory cells have been studied by immunohistochemistry in the lamina propria of the colon of children with Trichuris trichiura dysentery syndrome (TDS). No difference was found in the numbers of cells recognized by the monoclonal antibodies CD11c, CD68, or RFD7 between TDS children and local controls. However, large numbers of cells were recognized by an antibody against calprotectin (an anti-bacterial glycoprotein found in tissue infiltrating-monocytes) in TDS colonic mucosa, but few in control colon. Large numbers of cells containing tumour necrosis factor alpha (TNF alpha) were also seen in TDS mucosa; cells isolated from TDS mucosa secreted more TNF alpha than cells from control mucosa; and children with TDS had high levels of circulating TNF alpha. Non-specific macrophage-mediated inflammation and local cytokine production may therefore play a role in the pathogenesis of TDS.

The relationship between the lymph follicle and expression of HLA-DR antigens on the colonic epithelium was investigated. Colonic specimens containing lymph follicles were examined immunohistochemically for HLA-DR expression. Thirty-six biopsy specimens (29 from macroscopically uninvolved areas and seven from aphthoid lesions) were obtained from 10 patients with Crohn's disease, and another 43 specimens were obtained from 33 patients with various intestinal diseases (non-Crohn's disease). HLA-DR antigens were expressed on the epithelium around the lymph follicle in both groups. However, the rate of expression (31/36) in Crohn's disease was significantly higher than that (5/43) in non-Crohn's diseases (P < 0.001). In addition, the extent and intensity of expression were broader and stronger in Crohn's disease than in non-Crohn's disease. This study is the first demonstration of anomalous HLA-DR expression on the epithelium around the lymph follicle in Crohn's disease.

The effector function of immunocompetent cells in the gut mucosa has not yet been defined. The cytotoxic function of these cells might be important in the normal immune response and could be relevant to the mucosal damage seen in inflammatory conditions. The cytotoxic function of isolated intraepithelial and lamina propria mononuclear cells in six and 18 hour assays after the addition of various stimuli that interact with the human leukocyte antigens CD2 and CD3 on the mucosal effector cells was investigated. T cell phenotypes were determined using CD4, CD8, and HML1 to characterise cells of the appropriate compartments. Anti-CD3 and phytohaemagglutinin can induce toxic activity of lamina propria lymphocytes in most individuals after six hours and in all individuals after 18 hours. Anti-CD2, anti-CD3, and phytohaemagglutinin are similarly effective at triggering lamina propria lymphocytes. Intraepithelial lymphocytes contain predominantly CD8 and HML1 positive T cells, differentiating phenotypically intraepithelial lymphocytes from lamina propria lymphocytes. Intraepithelial lymphocytes are not cytotoxic at six hours, but have a toxic function comparable with lamina propria lymphocytes after 18 hours with all three triggers. Intraepithelial lymphocytes from inflamed mucosa (Crohn's disease and diverticulitis) mediate significantly reduced cytotoxicity in vitro compared with normal mucosa, whereas lamina propria lymphocyte toxicity is not different. Reduced numbers of cytotoxic cells and reduced reactivity to the trigger substances used after in vivo activation or cold target inhibition could explain the observed differences between intraepithelial lymphocytes from inflamed and uninflamed mucosa. Changes in cell mediated cytotoxicity of intraepithelial lymphocytes and lamina propria lymphocytes may be involved in the mucosal damage in these inflammatory conditions.

A six-month old baby, who had been fed for 2 months with casein hydrolysate formula (MA-1) for treatment of milk allergy, developed diarrhea. The baby's RAST score for milk was negative. IgE antibodies to MA-1 but not to MA-1 depleted of lipid (fat-free MA-1), were demonstrated by enzyme-linked immunosorbent assay (ELISA). It was confirmed by the appearance of numerous mast cells and eosinophils in a rectal smear taken after challenge with MA-1 (but not detected after challenge with fat-free MA-1) that some component in the lipid of MA-1 must be an allergen in this patient.

The immunoglobulins level, were estimated in the sera of 51 patients with different colonic disorders and 12 controls. In 27 of them, tissue immunoglobulin level were estimated. In bilharzial patients there was significant increase in the serum level of IgG, IgM and IgE. IgA and IgD showed no change. IgA containing cells were (87.5%), IgG (50%) and IgM (16.7%). In patients with amoebic colitis, there was significant increase in serum IgG and IgE. IgA and IgD showed significant decrease while IgM was within normal limits. Tissue IgA and IgG were detected in all acses. IgM containing cells were detected in 2 cases. In patients with irritable bowel syndrome (I.B.S.), there was significant high levels of IgM and IgE. IgG showed significant low level, while IgG and IgA showed no change. Tissue IgA were detected in (70%), IgG in (10%) and IgM in (20%). In patients with ulcerative colitis (U.C.), there was significant high levels of IgM and IgE. IgD showed significant low level, while IgG and IgA showed no change. Tissue IgA, IgG and IgM were detected in all cases. In patients with Crohn's disease, the 3 immunoglobulins were detected.

A complex clinico-immunological examination of patients subjected to extensive resections of different portions of the intestine was performed. It was established that participation of the immune system in regulation of homeostasis in patients under examination was characterized by lympho- and monocytopenia, lower amount of T-lymphocytes at the expense of low avidity fractions with a simultaneous increase of high avidity fractions of these cells.

In vitro immunoglobulin secretion has been studied using isolated colonic mucosal lymphoid cell populations obtained from 41 patients with non-inflammatory intestinal disease. Cell fractions were separated into intra-epithelial and lamina propria-enriched populations. The secretion of immunoglobulins by mucosal cells appeared to be independent of mitogen stimulation. When intra-epithelial lymphocyte preparations were co-cultured with autologous lamina propria lymphocytes, the secretion of IgM was significantly depressed but that of IgA and IgG was preserved. Co-cultured of mucosal cells with autologous peripheral blood lymphocytes resulted in suppression of pokeweed mitogen stimulated and unstimulated immunoglobulin secretion. Lamina propria T cells were shown to provide helper function for the in vitro secretion of IgA and IgM but not IgG, by autologous peripheral blood B cells. Immunoglobulin secretion by lamina propria lymphocytes was shown to be partly dependent on the concentration of E-rosette forming cells. These experiments demonstrate that human colonic mucosal lymphoid cells contain specific populations of helper and suppressor cells which selectively control intestinal immunoglobulin secretion.

Eighty four colonic biopsy specimens were obtained from patients with ulcerative colitis, Crohn's disease, radiation colitis, infectious colitis, and from normal controls. Paired specimens were examined by histological and immunohistochemical methods using monoclonal antibodies to the beta chain of HLA-DR antigen. The expression of HLA-DR antigen in mucosal epithelial cells was strongly related to whether the specimens were actively inflamed: epithelial cells from 34 of 37 inflamed specimens (nu three of 42 non-inflamed specimens) were HLA-DR positive (p less than 0.0001). Epithelial cells were uniformly HLA-DR negative in specimens from normal control patients despite the presence of HLA-DR positive lymphoid cells and macrophages in the lamina propria. Epithelial cells in specimens from patients with ulcerative colitis, Crohn's disease, and radiation colitis were HLA-DR positive in 30 of 33 inflamed biopsy specimens and in only three of 25 non-inflamed specimens (p less than 0.0001). Epithelial cells were HLA-DR positive in nine of 10 biopsy specimens from patients with acute infectious colitis (p less than 0.01).

The immunoreactivity of patients with colorectal or pancreatic cancer was investigated by a modified leukocyte adherence inhibition test (LAI). The microcapillary-LAI-test was easy to practise and measurements had a low intra-assay coefficient of variation. The number of false positive results was low, only 1 of 39 patients with benign disease and not one of 13 healthy volunteers reacted in the presence of colorectal carcinoma extracts. In 43 patients with colorectal cancer 17 of 25 (68%) with Dukes' B & C and 15 of 18 (83%) with Dukes' D cancer were LAI positive. Of the 19 patients with colon polyps 6 positive results were observed, exclusively in case of large villous adenomas, polyposis coli and one polyp with a focal carcinoma. 10 of 11 patients with pancreatic cancer reacted in the test. Leukocytes from patients with limited cancer of other source, when tested with colorectal or pancreatic carcinoma extracts, showed a negative LAI response. However, in case of metastatic diseases some crossreactions were observed, which resulted in false positive tests. Following our results, the LAI test could be of value in the diagnosis of gastrointestinal carcinoma.

The mesenteric lymph nodes in total colectomy specimens from patients with ulcerative colitis or Crohn's colitis were assessed for immunological reactivity. Despite the suggestion of previous work using different methods that immunological difference between the diseases exist and may be important in pathogenesis, no significant differences were found in this study. Changes were observed in both diseases and when ulceration was present these consisted of both cellular and humoral immune reactivity. There was no evidence of diminished cell-mediated immunity in Crohn's disease and the presence of granulomata did not constitute any special immunological group.

The survival of cells from five different cultures of allogeneic malignant colonic carcinoma, two from normal adult colonic epithelium and eight from foetal colonic epithelium in the presence of leucocytes from patients with neoplastic and inflammatory disorders of the colon has been compared. Cytotoxicity assessed by the reduction of the number of adherent target cells in microplate wells compared with those surviving in wells treated with tissue culture medium alone was observed with leucocytes from donors in all categories examined including those from individuals without any known abnormality. Patients with ulcerative colitis were the only group to reveal consistent reactivity against cultures derived from all three sources, an observation which may reflect sensitization to organ-related antigens in this disease. In contrast, leucocytes from patients with bowel neoplasia showed reactivity for cells derived from colon carcinoma tissue, which was comparable to that of healthy donors. Evidence for tumour-specific cytotoxicity was therefore lacking in this study. It is suggested that the detection of tumour-associated antigens on cultured cells may be limited by a number of factors of which the wide variation in reactivity among controls and unspecified nature of the target cells are likely to be of greatest importance.

Autoantibodies reacting with endocrine cells in the gastrointestinal mucosa were found by indirect immunofluorescence in 22 out of 268 sera (8.2%) obtained from patients with coeliac disease, Crohn's disease, ulcerative colitis, irritable bowel syndrome, and from subjects without bowel disease. A double immunofluorescence technique showed that the autoantibodies reacted with cells secreting gastric inhibitory polypeptide (glucose dependent insulinotropic polypeptide, GIP), secretin, somatostatin or enteroglucagon. Most sera contained antibodies against more than one cell type. Neither the presence of a particular antibody nor the pattern of antibody combinations appeared to be specific for any diagnostic category. The mean plasma GIP concentrations, however, both fasting and two hours after a test meal, were significantly lower in subjects with GIP cell autoantibodies. Thus gut hormone cell autoantibodies may be markers of impaired hormone secretion.

We isolated lymphocytes from the lamina propria of colon from 19 patients with inflammatory bowel disease, colon cancer, and certain benign conditions to determine: (1) if these lymphocytes could mediate mitogen-induced (MICC) and spontaneous cell-mediated cytotoxicity (SCMC), and (2) if there were any differences in cytotoxic effectiveness which could relate to the underlying disease. We found that lamina propria lymphocytes functioned well in MICC reactions with phytohaemagglutinin, but not concanavalin A as the inducing mitogen (specific lysis 28 5% vs 5 3%). Lamina propria lymphocytes did not mediate SCMC (specific lysis 0.3%). Neither the presence of inflammation not the underlying disease of the patient influenced the cytotoxic activity. Peripheral blood lymphocytes from normal subjects and patients performed well in MICC assay with both phytohaemagglutinin and concanvalin A as the inducing mitogen and were equally effective in SCMC reactions.

Two hundred and thirty-seven patients with carcinoma of the colon and 16 patients with benign lesions of the colon and rectum underwent skin tests with 2-4, dinitrochlorobenzene and a battery of intradermal antigens. The incidence of 2-4, dinitrochlorobenzene reactors decreased with the increasing stage of the disease. Seventy-six per cent of the patients with Dukes' A cancer were 2-4, dinitrochlorobenzene positive compared with 56 per cent of those with Dukes' B cancer and 61 per cent of those with Dukes' C lesions. Of the patients with advanced primary operable cancer, those who have metastases beyond the intestine and its mesentery, only 46 per cent were 2-4, dinitrochlorobenzene positive. Only 42 per cent of the patients with inoperable advanced or recurrent disease reacted to 2-4, dinitrochlorobenzene. Neither age nor sex was a determinate factor in the capacity of the patient to respond to 2-4, dinitrochlorobenzene. Tumor burden appeared to correlate best with the ability of the patient to respond to 2-4, dinitrochlorobenzene. In patients with Dukes' A or B lesions, the clinical follow-up period was too short to gauge prognostic significance of skin tests. In patients with Dukes' C lesions who were observed at 12 months, six of 11 in the 2-4, dinitrochlorobenzene negative group had a recurrence or died of disease compared with only four of 17 in the 2-4, dinitrochlorobenzene positive group, p less than 0.10. In 38 patients with advanced primary operable cancer who were observed for nine months, 40 per cared iwth 28 per cent of 2-4, dinitrochlorobenzene positive group. A similar relationship was observed in a group of patients with advanced or recurrent disease who were observed for nine months in which 58 per cent of the patients in the 2-4, dinitrochlorobenzene negative group were dead of disease compared with 40 per cent of those in the 2-4, dinitrochlorobenzene positive group. Skin testing with 2-4, dinitrochlorobenzene and selected intradermal antigens adds prognostic information to that predicted from the clinicopathologic stage of the disease in instances of carcinoma of the colon and rectum. In general, patients with reactive skin tests have more favorable recurrence and survival rates with each stage of the disease.

Recently, the demonstration of serum agglutinins to Eubacterium and Peptostreptococcus strains has been found to be useful as a diagnostic test for Crohn's disease. Therefore, conditions determining the occurrence of these antibodies were studied in patients with Crohn's disease and ulcerative colitis. Localization of Crohn's disease in the colon, the presence of fistulae and serum levels of immunoglobulins were found to be contributory determinants for the occurrence of the agglutinins.

A significant association between carcinoembryonic antigen (CEA) assay titers and peripheral lymphocyte counts was observed in 148 simultaneous determinations. The association was present in a high cancer risk study group of patients with myasthenia gravis and in a control group of patients with granulomatous disease of the bowel, ulcerative colitis, and colonic neoplasms. A highly significant difference in the percentage of positive CEA assays in relation to lymphocyte counts was noted both in the study and control groups. In the study group CEA was positive in seven of 41 patients (17 percent) with counts above 2,000 per cubic millimeter, and in 26 of 56 (43 percent) of those with lower counts (X2 = 9.06, p smaller than 0.005). The corresponding percentages in the control group were 20 and 61 percent (X2 = 5.60, p smaller than 0.025). A significant difference between the means of lymphocytes in groups with different CEA titers also was found (F = 6.77, p smaller than 0.05). The finding of lower peripheral lymphocytes and/or higher titers of CEA in groups with increased risk of cancer, i.e., severe myasthenia gravis, patients with thymomas and patients with long history of granulomatous disease of the bowel, suggests on association between the results of the two tests and cancer risk. The observations indicate that the yield of CEA assay in cancer detection may be higher in patients with lwo lymphocytes, and that these determinations used in conjunction may serve better as diagnostic and prognostic aids in patients with known neoplasms or in high cancer risk groups.

The serum levels of CA 19-9 and carcinoembryonic antigen (CEA) were determined in 37 patients with benign colorectal diseases and in 111 patients with newly discovered colorectal carcinomas or clinically verified relapses. In cancer patients, the CA 19-9 level ranged from normal (0-37 U ml-1) to 77,500 U ml-1 whereas all samples but one from patients with benign colorectal diseases had a normal value. CA 19-9 was increased in 46% and 45% of patients with an advanced (Dukes C or D) carcinoma or a verified recidive, respectively. Only one out of 26 patients (4%) with a localized (Dukes A or B) carcinoma displayed an elevated CA 19-9 level (greater than 37 U ml-1). No clear correlation was found between the CA 19-9 and CEA levels. The sensitivity of the CA 19-9 test (36%) was poorer than that of the CEA assay (69%), but the new test was markedly more specific (97% vs 70%) than the CEA assay.

In Brown-Norway (BN) rats, oral administration of mercuric chloride (HgCl2) induced morphological lesions of the ileum and, in lesser degree, of the colon, with abnormal deposits of IgA in the basement membranes of intestinal glands and of IgG in the basement membranes and in the lamina propria. IgG reactive with renal and intestinal basement membranes and in the lamina propria. IgG reactive with renal and intestinal basement membranes and with the lamina propria of a normal BN rat was found in the serum and IgG deposits were present in renal glomeruli of BN rats receiving HgCl2. Thus, it is conceivable that the deposits of IgG present in the intestine resulted from local fixation of circulating autoantibodies. In contrast, IgA with basement membrane reactivity was not detected in the sera nor in the renal glomeruli, suggesting that the intestinal deposits of IgA were formed in situ. This IgA-IgG intestinal disease inducible in BN rats may provide a model for the study of alterations of the secretory IgA system, as well as for testing the possibility that abnormal deposits IgA-IgG in the intestinal structures are associated with local functional changes.

Lamina proprial lymphocytes (LPL), isolated by an EDTA-collagenase technique from patients with various colonic diseases, were investigated for LMIF release in vitro. On stimulation with the preparation of Kunin antigen, macrophage-depleted LPL from patients with severely or moderately active ulcerative colitis showed LMIF release which was significantly greater than that observed using LPL from patients with mild colitis or from those with other diseases of the large bowel, including Crohn's disease. Results similar to those obtained with LPL were found with the corresponding peripheral blood lymphocytes (PBL) stimulated by the preparation of Kunin antigen. In contrast, nonspecific stimulation in vitro with Concanavalin A showed no differences in LMIF releases by the LPL or PBL in the various disease groups. It is suggested that hypersensitivity to Kunin antigen may have pathogenic significance in ulcerative colitis.

The tube leukocyte adherence inhibition (LAI) technique was used to investigate the antitumor immunity in two groups of patients generally considered to be at "high risk" of developing colorectal cancer. The first group comprised 21 patients with colorectal polyps and the second 12 patients with various forms of colitis. Also 29 patients with histologically confirmed colorectal cancer were tested. The tube LAI assay was performed using peripheral blood leukocytes from individual patients and crude extracts of colorectal and breast cancers. Positive LAI reactions were observed in 18 out of 29 (62%) patients with colorectal cancer, in 1 out of 21 (5%) patients with colorectal polyps and in 1 out of 12 (8%) patients with colitis. The results indicate that in confirmed cases of malignancies, sensitization to colon tumor-associated antigens could be detected in the tube LAI test, whereas, premalignant sensitization to these antigens in "high risk" groups of patients could not be demonstrated.

A 43-year-old male developed seronegative polyarthritis after a short attack of diarrhoea. For about two years before he had experienced intermittent abdominal pain, distension and constipation. Laparotomy showed a chronic sigmoid volvulus for which sigmoid colectomy was performed. Post-operatively he had no further bowel symptoms, no further arthritis and there was radiological improvement of involved joints. Tissue typing showed HL-A B27 antigen.

Monoclonal antibody (MAb) B72.3 was generated against a carcinoma metastasis and has been shown to bind with a high degree of selectivity to a tumor-associated glycoprotein (TAG-72) found in human colon and breast carcinomas, while showing minimal reactivity to any normal adult tissues. Competition radioimmunoassays have been developed for the detection of TAG-72 in tumors and sera from both athymic mice bearing human carcinoma xenografts and patients with colon, breast and other carcinomas. The distribution of TAG-72 in human tumor xenografts was restricted to tumors originating from the LS-174T human colon carcinoma, with no significant reactivity being detected in human tumor xenografts from a different colon carcinoma, a human breast carcinoma, or a human melanoma. The levels of TAG-72 in clinical material obtained from surgery were examined; high levels were found in colon carcinomas and to a lesser extent in breast carcinomas, while no detectable levels were found in normal tissues. Sera from apparently normal patients contained a mean level of 2.2 units per ml of TAG-72. A level of 3 standard deviations above the mean level of TAG-72 found in normals was employed as a cut-off value to indicate a positive test result in subsequent studies. No patient with inflammatory disease or other benign colon disease exhibited elevated levels of TAG-72. Seven out of 20 patients with other carcinomas had elevated serum levels of TAG-72. The serum TAG-72 levels were compared with the serum levels of antigens recognized by the MAbs currently used to screen sera of carcinoma patients (CEA, GICA, and OC125). It was clearly demonstrated that TAG-72 is different from these antigens and can be found in some sera in which no antigen is detected by otherwise available MAb RIAs.

Food allergy in childhood presents with different clinical manifestations depending on the age of the affected child. Whereas toddlers and children with food allergy or pseudo-allergy present with similar symptoms as adults, two different forms have been identified in the newborn and infant period. One can occur as allergic colitis in breast or bottle fed infants. In breast fed infants the bloody mucoid stools are produced because of an allergic reaction of the colonic mucosa to foreign proteins which are delivered to the baby via breast milk. In bottle fed infants the given foreign protein itself can trigger the allergic reaction. The clinical, endoscopical and histological appearance is similar to that of ulcerative colitis. Elimination of foreign proteins from the diet of the mother or in bottle fed infants from the child is the therapy of choice. The second manifestation of food allergy in this age group is the cow's milk protein intolerance with predominantly gastrointestinal symptoms such as vomiting, diarrhea and failure to thrive. The diagnosis is based on the clinical picture alone. The usual laboratory tests don't discriminate enough and can therefore not confirm the diagnosis. Elimination of the affecting protein and replacement by a semi-elementary diet are recommended for therapy. The prevention of allergies by dietetic means has become of great importance since it was possible to identify newborns at risk for allergies. The prolonged breast feeding and the late introduction of solids later than the sixth month of life is the preventive measure. "Hypoallergenic" formulae are not recommended because not enough solid data are available to confirm their preventive effect.

In this chapter we have outlined the seronegative spondarthritides associated with bowel disease, excluding those discussed in separate chapters. Although, traditionally, Crohn's disease and ulcerative colitis have been dealt with separately in any discussion of seronegative arthritides, they have been discussed together here for the following reasons. Despite being pathologically distinct they show remarkable similarity in extraintestinal manifestations including any associated arthritis. Any observed differences in prevalence rates of arthritis may be secondary to the relative difficulty in diagnosing Crohn's disease, and most of the important prevalence studies were done before the more sophisticated techniques to investigate the bowel became available. This may in part explain the dramatic increase in the world-wide incidence of Crohn's disease seen in the last 30 years, particularly through the 1970s, but which has fallen off recently (Miller et al, 1974). It would seem that the body has a limited means of expressing disease processes. The final common pathway of a number of quite distinct disease entities is the concept of the seronegative spondarthritides. Exactly how the microbiological, immunogenetic and molecular factors interact to produce a particular disease end-point is currently not clearly defined, but with the increasingly sophisticated means to investigate the body at a cellular level the explanations may soon be at hand. Further controlled family studies are also needed to define the genetic relationships more precisely. We may then be able to piece the jigsaw puzzle together.

The pathogenetic and clinical importance of intestinal spirochaetes in man is still unresolved. In 12 patients mainly presenting with mild diarrhoea, light and electron microscopy demonstrated massive spirochaetal infestation of the colonic mucosa (spirochaetosis). There were several hitherto unreported features: spirochaetes adhered not only to the surface epithelium of the intestine but were also present within epithelial cells and subepithelial macrophages; many partially degranulated mast cells were noted within the epithelium; there was a marked increase of IgE plasma cells within the lamina propria. In control biopsies intraepithelial mast cells were absent and IgE cells occurred only sporadically. Penetration of the microorganisms into the intestinal mucosa may be responsible for this unusual immune response. Spirochaetes, symptoms and findings disappeared after antibiotic therapy. The authors therefore suggest that intestinal spirochaetosis can cause clinical symptoms in man, and that spirochaetes should not invariably be considered harmless commensals.

The distribution patterns of epithelial blood group AB antigens (BG) in colon polyps of varying degrees of differentiation were studied by the mixed cell agglutination reaction. BG appeared in colon polyps if a certain degree of dedifferentiation was present. Two different distribution patterns were recognized: 1) association of BG with the secretory part of goblet cells with slight-to-moderate atypia and 2) association of BG with the whole cells in cases of pronounced dedifferentiation and loss of secretory activity. The second type resembled the distribution pattern of BG found previously in colon carcinomas. With respect to BG, the mucosa of colon polyps behaved similarly to embryonal colon mucosa. The detection of BG represents a useful method to assess objectively the degree of dedifferentiation in most colon polyps.

We studied the class-specific antibody response to the cow's milk antigen beta-lactoglobulin (beta-LG) in sera from patients with ulcerative colitis and Crohn's disease. IgG and IgM to beta-LG were significantly higher in patients when compared to healthy non-atopic controls, whereas IgA values were similar, and specific IgE absent in all groups. No correlation between IgG- or IgM-containing immune complexes was found with the corresponding isotype of antibody to beta-LG; however, IgM complexes correlated with serum total IgM in ulcerative colitis. In these patients, IgG antibodies were higher in active cases, whereas IgM increased in patients without signs of disease activity. Antibody titers did not correlate with disease duration or administration of antiinflammatory drugs. This pattern of anti-beta-LG reactivity suggests that the presence of intestinal lesions may be revealed by the selective increase of some antibody isotypes to orally administered antigens. Enhanced mucosal permeability may be studied by this type of serological analysis.

We investigated the pattern of genetic susceptibility to rheumatic diseases in Indians by means of HLA analysis. HLA B27 was present in more than 90% of cases included under the broad category or seronegative spondarthritides. In this respect our data resembled results reported for Caucasian populations. In our population, however, the phenotypic frequency of HLA B27 was low as reported from Japan.

Using an enzymatic method for their isolation, it has been shown that human colonic lamina proprial lymphocytes, isolated from patients with various colonic diseases and depleted of polymorphonuclear leukocytes and macrophages, demonstrated K-cell activity in two antibody-dependent cell-mediated cytotoxicity assays in vitro. No differences were found between the activities of lamina proprial lymphocytes isolated from colonic specimens involved by Crohn's disease, chronic ulcerative colitis, or colonorectal carcinoma. K-cell activity was demonstrable using lymphoid cells isolated from the colons of patients receiving steroid therapy at the time of the therapeutic resections. These findings indicate the need for further investigation of possible roles for K-cells in local immune responses in the human bowel mucosa in inflammatory and neoplastic diseases.