Studies on aziridine derivatives. III. Synthesis and immunopharmacological activity of aziridine derivatives of propionic acid

Several new aziridine derivatives of propionic acid were synthesized (10-15, 19, 20). o-, m-, p-Chloroanilide of chloroacetic acid 1-3 and chloride of 3-/p-chlorobenzoyl/acrylic acid 16 were the substrates. The compounds 1-3 in reaction with nicotine aldehyde or p-chlorobenzaldehyde were transformed into appropriate anilides of 2,3-epoxypropionic acid 4-9. These, in turn reacted with ethylenimine giving the appropriate 3-aziridine derivatives 10-15. Acid chloride 16 in reaction with amines gave the appropriate amides 17 and 18 which formed 2-aziridine derivatives 19 and 20 when under the influence of ethylenimine. Pharmacological analysis revealed that the aziridine derivatives 12-15, 19 and 20 modulate some immunological reactions with the prevailing effect of the suppressive component (PFC, RFC, IgM level, cellular response to SRBC). The stimulatory effect was observed with some compounds on the level of circulating IgG and GvH reaction. The mechanism of these compounds consists in their interference with the activity of Ts cells and mediators of the immunological reactions.     

The histogenesis of inflammatory fibroid polyps of the gastrointestinal tract

Inflammatory fibroid polyps are lesions occurring in the submucosa of the gastrointestinal tract. These lesions have been identified by a variety of names, indicative of their uncertain histogenesis. Three cases were studied by light microscopy and cytochemistry and, in one case, by electronmicroscopy. The most characteristic feature consisted of concentric fibrovascular structures. Peroxidase reaction for muscle actin and Ulex europeus agglutinin was positive in the two main cellular components. Electronmicroscopy revealed that the two principal cell constituents were endothelial and myocytic cells. These observations support the conclusion that inflammatory fibroid polyps are lesions of vascular origin.     

Characterization and quantification of cellular infiltrates in nasal mucosa of patients with grass pollen allergy, non-allergic patients with nasal polyps and controls.

Little is known about cellular infiltrates in nasal mucosa and the differences between these infiltrates in allergic and non-allergic patients. A reproducible and objective method making use of monoclonal antibodies for the quantification and characterization of cellular infiltrates in biopsy specimens of nasal mucosa is described. This method was used to study quantitative differences in cellular infiltrates in the epithelium and lamina propria of the nasal mucosa of patients with isolated grass pollen allergy, non-allergic patients with nasal polyps, and controls. A surprisingly wide variation was found in all groups. In all groups the T lymphocytes were much more numerous than the B lymphocytes. The number of CD8+ cells exceeded the number of CD4+ cells in the epithelium but in the lamina propria the numbers were approximately equal. Significant differences between the three groups were found with respect to the number of CD1+, IgE+, neutrophils and cytoplasmic IgG4+ cells. No significant differences were found in the numbers of CD4+, CD8+, CD14+, CD22+, HLA-DR+, IgG1-3+ cells or eosinophils. The use of biopsy in combination with monoclonal antibodies is an easy and well-tolerated method to study immunological reactions in the nasal mucosa. The results of this study indicate a possible role for a T-cell-mediated response in allergic rhinitis.     

Histamine release from human colonic mucosa in response to anti-IgE

Testing of tissue particles for mediator release may be very useful for the diagnosis of localized immunological abnormalities or allergies. The aim of this study was to set up a general procedure to test the reaction of large bowel mucosa to stimuli via the IgE-mediated pathway. Therefore, tissue particles from normal subjects and from patients suffering from different diseases (Crohn's disease, ulcerative colitis, intestinal polyps) obtained at routine coloscopy were exposed to either Hanks or anti-IgE solution to determine the spontaneous or the anti-IgE-induced histamine release, expressed as the percentage of the total histamine content of the biopsy. Histamine was measured using the single isotope radioenzymatic assay.In general, whereas anti-IgE interestingly reduced the histamine release compared to the spontaneous in most of the patients within the polyps group, there was a stimulating effect of anti-IgE throughout all other groups. Thus, the study confirms the possibility of performing functional tests using biopsy particles from the colon.     

Cellular differentiation status of epithelial polyps of the colorectum: the gastric foveolar cell-type in hyperplastic polyps

AIMS: The 'metaplastic' polyp of the colorectum, a synonym for the hyperplastic polyp, was named based only on features of the crypt epithelium. It is considered non-neoplastic, but the precise cellular differentiation status remains to be proven. METHODS AND RESULTS: Forty-eight hyperplastic polyps, 12 serrated adenomas, 45 tubular adenomas and five juvenile polyps were studied for their phenotypic expression using gastric (foveolar or pyloric gland cell), small intestinal (goblet cell), and colonic (goblet cell) cellular markers by immunohistochemical and mucin histochemical techniques. Gastric foveolar cell-type differentiation was significantly expressed in hyperplastic polyps, while colonic differentiation was also consistently preserved. Neither gastric pyloric-type nor small intestinal differentiation was observed. The same cell differentiation status as hyperplastic polyps was observed in serrated adenomas but not in tubular adenomas or juvenile polyps. CONCLUSIONS: A large proportion of hyperplastic polyps are composed of hybrid epithelium, with bidirectional differentiation to both gastric foveolar and colonic epithelial cells in the same crypt. Therefore hyperplastic polyps might be interpreted as the outcome of abnormal cell differentiation of stem cells. The same phenotypic expression suggests that hyperplastic polyps and serrated adenomas share the same cell lineage.     

Cross reaction between proteins from Larrea divaricata Cav. (jarilla) and cellular and extracellular proteins of Pseudomonas aeruginosa

Larrea divaricata is widely used in folk medicine to treat different pathologies, but little is known about its immunological properties. Pseudomonas aeruginosa is an opportunistic pathogen which causes several intrahospitalary infections. We aimed to assess the immunological relation between proteins from a crude extract of L. divaricata Cav. (JPCE) and cellular and extracellular proteins (EP) of P. aeruginosa, as well as to establish the cross reactivity between proteins of both species using a mouse anti-JPCE serum. Protein profiles of JPCE and P. aeruginosa were analyzed by SDS-PAGE. The percentage of similarity of protein bands between these two species was 43-57%. However, JPCE proteins were immunogenic. The reactivity of mouse anti-JPCE antibodies against different fractions was studied by western blot. The anti-JPCE serum detected several antigenic bands on different bacterial proteins. Several common immunoreactive bands were detected (27-100%) when bacterial proteins were incubated with anti-JPCE serum (heterologous reaction) and anti-bacterial proteins serum (homologous reaction). By enzyme-linked immunosorbant assay (ELISA) assays, high titers of anti-JPCE against different types of cellular bacterial fractions were observed (1/1280-1/2080). Our data clearly demonstrate that antibodies elicited with L. divaricata crude extract are able to cross-react with cellular and EP of P.aeruginosa. These findings could be relevant in the development of alternatives therapies for patients suffering intrahospitalary opportunistic infections with P.aeruginosa.     

Suppressor adherent phagocytic cells in solid tumors: a postulated escape mechanism.

A localized graft-versus-host reaction was used to assess the cellular immunity of mice supporting progressive growth of an allogeneic tumor. It was found that early in tumor development anti-tumor-cellular-mediated immunity was hyper-responsive, and that adherent phagocytic cells isolated from the neoplasm would inhibit the specific immunological response. Based on these observations and indirect inference it was postulated that a major factor in the success of an immunogenic tumor is the coterminous development of a normal adherent phagocytic cell population which specifically nullifies, in situ, cellular-mediated anti-tumor immunity.     

Resorption of insoluble,heterologous, fluorescein-collagen sponges in sensitized and non-sensitized rats.

Sponges of insoluble bovine collagen were slowly resorbed over a 35-day period when implanted under the back skin of rats. The cellular picture was typical of a mild foreign-body reaction. The reaction to fluorescein-labelled collagen sponges was similar but there was evidence also of a weak immunological response. An acute inflammatory reaction with massive oedema was elicited when fluorescein-labelled collagen sponges were implanted in rats previously sensitized to either fluorescein-collagen or fluorescein-bovine serum albumin. The early invasion by PMN leucocytes subsided after 4 days and caused no observable breakdown of the sponge. The implanted material was rapidly encapsulated by fibrous tissue which was then resorbed along with the sponge between the 7th and 12th day. Macrophages were very active in the sponge at this time, sometimes forming giant cells. Fibroblasts were invading from the periphery with the development of the granulation tissue. The small residue which remained after this time was overrun by granulation tissue and was slowly resorbed up to the 35th day. Throughout the period of study there was only a weak local immunological response after the 28th day. The level of circulating antibodies against the fluorescein hapten was high, but the titre for the antibodies against bovine collagen remained low. The significance of these findings in the pathological destruction of connective tissue is discussed.     

Expression of a high molecular weight mucin-type glycoprotein in human colon cancer as defined by monoclonal antibody 1D3

Murine monoclonal antibody 1D3 recognizes a high molecular weight acidic mucin restricted to the epithelium of normal colonic mucosa and goblet cells. Of 72 colonic carcinoma specimens examined, 29 were found to have detectable level of 1D3 antigen by an indirect immunoperoxidase staining assay on fixed, paraffin-embedded tissue sections. In some specimens a focal staining pattern was observed, while in others 50-90% of tumor cells were stained. Of 28 cases having adjacent normal mucosa, all 28 showed intense staining reaction in the normal mucosa and goblet cells despite the fact that 18 of the tumors were unstained. One of 1 colonic diverticulosis, 2 of 2 ulcerative colitis, 3 of 3 villo-glandular polyps, 19 of 20 adenomatous polyps and 17 of 19 hyperplastic polyps were also stained heavily for the 1D3 antigen. Colonic carcinomas displayed a range of staining patterns and a great degree of antigenic heterogeneity. Well-differentiated tumors characterized by typical goblet cells were almost always positive (10 of 12). As cellular structure became disorganized, as in moderately-differentiated tumors, about 33% of the tumors (17 of 51) stained for 1D3 antigen. As the tumor became more invasive with further disorientation of cellular features, as in poorly differentiated tumors, very few specimens (2 of 9) were positive. It was apparent that with the progression of de-differentiation there was a gradual loss of 1D3 antigen in human colonic tumors.     

Cross reaction between proteins from Larrea divaricata Cav. (jarilla) and cellular and extracellular proteins of Pseudomonas aeruginosa

Larrea divaricata is widely used in folk medicine to treat different pathologies, but little is known about its immunological properties. Pseudomonas aeruginosa is an opportunistic pathogen which causes several intrahospitalary infections. We aimed to assess the immunological relation between proteins from a crude extract of L. divaricata Cav. (JPCE) and cellular and extracellular proteins (EP) of P. aeruginosa, as well as to establish the cross reactivity between proteins of both species using a mouse anti-JPCE serum. Protein profiles of JPCE and P. aeruginosa were analyzed by SDS-PAGE. The percentage of similarity of protein bands between these two species was 43–57%. However, JPCE proteins were immunogenic. The reactivity of mouse anti-JPCE antibodies against different fractions was studied by western blot. The anti-JPCE serum detected several antigenic bands on different bacterial proteins. Several common immunoreactive bands were detected (27–100%) when bacterial proteins were incubated with anti-JPCE serum (heterologous reaction) and anti-bacterial proteins serum (homologous reaction). By enzyme-linked immunosorbant assay (ELISA) assays, high titers of anti-JPCE against different types of cellular bacterial fractions were observed (1/1280–1/2080). Our data clearly demonstrate that antibodies elicited with L. divaricata crude extract are able to cross-react with cellular and EP of P.aeruginosa. These findings could be relevant in the development of alternatives therapies for patients suffering intrahospitalary opportunistic infections with P.aeruginosa.     

声带息肉的组织病理学

对150例声带息肉手术摘除标本进行组织病理学检查,结果显示息肉上皮下组织可见水肿、血管扩张、玻璃样变、纤维增生、灶性出血、含铁血黄素沉着等多种改变.炎症细胞浸润十分常见.结合文献复习,对本病的病理分型进行讨论.     

Glycoprotein abnormalities in colonic carcinomata, adenomata, and hyperplastic polyps shown by lectin peroxidase histochemistry.

A technique for lectin-peroxidase histochemistry was adapted for the study of formalin fixed paraffin embedded colonic tissue. Ten lectins with differing carbohydrate binding specificity were tested against 20 normal rectal biopsy specimens and tissue from 19 colonic carcinomata, 19 tubular or tubulovillous adenomata, and 19 hyperplastic polyps. None of the normal rectal biopsy specimens bound the lectins peanut agglutinin (PNA), Griffonia simplicifolia II (GSII), and Ulex europaeus I (UEAI), whereas 18 carcinomata, 12 adenomata, and 18 hyperplastic polyps showed affinity for one or more of these lectins. Hyperplastic colonic polyps are shown to possess similar abnormalities in glycoprotein structure to malignant and adenomatous colonic tissue. This may simply indicate a non-specific reaction to changed rates of cell proliferation but might represent a more fundamental association between hyperplastic polyps and adenocarcinomas.     

Failure of correlation between in vitro and in vivo effect of T and B cell mitogens in the mouse.

T and B cell mitogens as PHA, PWM, LPS, SIII, DS, PVP were investigated in vitro in direct MI assay with mouse PECs as well as in vivo for their phagocytosis-enhancing capacity. All mitogens induced a dose-dependent MI although of differing degrees. In contrast, a phagocytosis-stimulating effect in vivo could be observed only by using the mitogens of bacterial origin such as LPS and SIII; the other mitogens induced, however, rather a loss of phagocytosis, although a cellular reaction in the sense of an increased PEC number was partly registered. The reactivity of PEC to LPS but not to PHA disappeared in the MI assay after foregoing desensitization of mice with the corresponding mitogen, suggesting the existence of a cellular immunity against the bacterial antigen. The importance of the induction of cell-mediated events in infection immunity in a classical immunological way will be discussed.     

Colorectal polyps with extensive absorptive enterocyte differentiation: histologically distinct variant of hyperplastic polyps

BACKGROUND: The histologic classification of colorectal polyps is well established. However, practicing pathologists may still occasionally encounter colorectal polyps that are difficult to classify. We studied 6 colorectal polyps that showed uncommon histologic features that have not been described in the English language literature. MATERIALS AND METHODS: The polyps were studied using standard hematoxylin-eosin stain, mucin histochemistry, and electron microscopy. RESULTS: The 6 polyps we studied showed extensive papillary and villous structures with alternating villi and crypts. The villi were lined by well-differentiated absorptive cells, whereas the crypts were lined by immature glandular cells, thus mimicking the histology of the small intestinal mucosa. CONCLUSIONS: These polyps appear to represent a variant of the hyperplastic polyp, in as much as cellular maturation (immature glandular cells differentiate into the mature surface absorptive cells) is the essential feature distinguishing hyperplastic polyps from adenomas.     

Immunosuppressory mini-regions of HLA-DP and HLA-DR.

Our previous studies showed, that the,TPQRGDVYT, QRGDVYT and RGDVYT fragments, located in the beta164-172 loop of HLA-DQ, strongly suppress the humoral and cellular immune response, while their shorter analogs, RGDV, RGDVY, and QRGDVY, show only weak stimulatory activity in respect to humoral immunological response. The fragments contain the RGDVY sequence that is analogous to thymopentin (pentapeptide RKDVY, an immune system activator) as well as the RGD sequence, known for its importance for cellular association phenomena. Based on the crystal structure of HLA-DR1, we also designed and synthesized a cyclic analog C*RGDVYC* (where C* indicates Cys participating in disulfide bridge) with restricted conformation, which strongly suppresses both humoral and cellular immune response. In the present study we synthesized and tested the immunological properties of the linear and cyclic HLA-DP and HLA-DR counterparts of all the above HLA-DQ fragments. Although the results show that the linear HLA-DP fragments possess moderate immunosuppressory potency, their conformationally restricted analog, C*QGDVYC*C shows a considerable suppression of both humoral and cellular immune response. The nonapeptide fragment of HLA-DR, VPRSGEVYT and particularly its cyclic analog C*SGEVYC*, are strong suppressors of the humoral response.     

Cell-mediated immunity to varicella-zoster virus measured by virus inactivation: mechanism and blocking of the reaction by specific antibody.

The process whereby varicella-zoster (V-Z) virus is inactivated in vitro by immune human peripheral blood leukocytes stimulated with V-Z antigen was examined. It was found that stimulation of leukocytes by V-Z antigen, but not by other viral antigens, was required for inactivation of V-Z virus to occur. Viral inactivation could be blocked by addition of V-Z antiserum to either the stimulation phase of the reaction or the inactivation phase, further demonstrating the specificity of the reaction. In addition these blocking experiments suggested that modulation of V-Z membrane antigen by antiserum occurred with an accompanying loss of immunological recognition of virus-infected cells. Inactivation of V-Z virus in vitro in this study appeared not to be dependent upon the secretion of interferon or upon antibody-dependent cellular cytotoxicity. The specific cells required for V-Z inactivation were T lymphocytes and monocytes (macrophage precursors).     

Immune potential in human uraemia. 2. Changes after regular haemodialysis therapy.

Parameters of both humoral and cellular immune potential were measured in a group of patients with severe renal failure before and after three months' regular haemodialysis therapy. Evidence is presented of improvement in cellular immune potential and of a tendency of the response of lymphocytes to PHA to return to normal. No improvement in humoral responsiveness was demonstrable, and it is suggested that uraemic patients on regular haemodialysis may have an impaired capacity to establish new immunological memory.     

Immunolocalization of beta catenin in intestinal polyps of Peutz-Jeghers and juvenile polyposis syndromes

AIM: To examine the membranous and nuclear distribution of beta catenin in the epithelial cells of gut polyps from Peutz-Jeghers syndrome and juvenile polyposis in comparison with other types of polyps and tumours. METHODS: Immunohistochemistry for beta catenin and proliferation markers was performed on conventional paraffin sections. Immunohistological staining was carried out on Peutz-Jeghers syndrome polyps from four different families, on juvenile polyposis polyps from two different families, on solitary juvenile polyps, and on hyperplastic polyps. The immunohistochemistry was evaluated qualitatively in relation to defined areas of the polyps. RESULTS: All polyps from the hamartomatous polyposis syndromes (Peutz-Jeghers syndrome and juvenile polyposis) showed nuclear localization of beta catenin in some epithelial cell nuclei. In Peutz-Jeghers syndrome polyps beta catenin positive nuclei were seen at the base of the deep crypt infoldings. In juvenile polyposis polyps and in some solitary juvenile polyps they were found in irregularly distributed cryptal epithelial cells corresponding to the proliferative compartments. Normal mucosa of the gut and hyperplastic polyps of the colon do not show nuclear staining for beta catenin. CONCLUSIONS: The dysregulation of cellular beta catenin distribution is not only a phenomenon of adenoma formation and adenoma progression in the colon--it is at least focally present in polyps of the hamartomatous type and is related to the proliferation zones of these polyps. The nuclear translocation of beta catenin most probably reflects a disturbed beta catenin metabolism. In view of the different functions of beta catenin during development and cell differentiation, the nuclear translocation of beta catenin is likely to be an important factor in enhanced cell proliferation which escapes local control mechanisms.     

Sessile adenoma of the gallbladder. Reappraisal of its importance as a precancerous lesion

Three hundred gallbladders from patients with cholelithiasis were examined under a dissection microscope. Sixteen (5%) were found to have what have been called sessile adenomas. They consisted of mixtures of hyperplastic lining epithelium and metaplastic mucous glands, and their interstitium often included smooth-muscle fibers. Small foci of moderately severe cellular atypism were present in 19% of the adenoma cases, but none had definitive evidence of malignancy. Carcinoembryonic antigen was demonstrated in hyperplastic lining epithelium with or without cellular atypism in 31% of the cases. Sessile adenomas most likely represent reactive overgrowth and therefore we prefer to term them hyperplastic polyps.