Lectin histochemistry of normal bronchopulmonary tissues and common forms of bronchogenic carcinoma

Normal bronchopulmonary tissues, areas of basal cell hyperplasia and of squamous metaplasia, and 26 pulmonary carcinomas, including the three major types (squamous cell carcinoma, adenocarcinoma, and oat cell carcinoma), were studied with 12 biotinylated plant lectins. The study demonstrated a rather characteristic lectin-binding profile for most normal epithelial cell types, while pulmonary tumors showed a variation in the lectin reactivity when compared with the whole spectrum of normal cells, with binding of some lectins that were not reactive with normal tissues and a loss of reactivity with others that bound to normal tissues. Adenocarcinomas showed the highest density of reacting sites, while undifferentiated carcinomas were the less-reacting variant. Areas of metaplasia and hyperplasia showed some variation in the lectin-binding profile with normal tissues, but as a whole, there was a much smaller number of reacting sites than tumors.     

Recombinant forms of Gerbich blood group antigens: expression and purification.

Recombinant forms of normal glycophorin C (GPC), carrying the high frequency Gerbich blood group antigens, and its natural deletion mutants of Yus and Ge type (all combined with oligohistidyl tag) were expressed in CHO and COS 7 cells. The stable expression of all recombinant forms of GPC in CHO cells was obtained, but the level of expression was low and detectable only by flow cytometry. The high level of transient expression of GPC recombinant forms in COS 7 cells allowed their purification on Ni-NTA-agarose. The purified recombinant GPC and mutants of Yus and Ge type behaved in SDS-PAGE similarly to normal GPC forms from RBC membranes. The recombinant GPC.Yus and GPC.Ge mutants appeared as diffuse bands, suggesting the similar heterogeneity of glycosylation that was observed in natural GPC.Yus and GPC.Ge glycoproteins. The flow cytometry analysis of the transfected CHO and COS 7 cells showed that binding of anti-GPC monoclonal antibodies to GPC variants was accordant with the known fine specificity of these antibodies. The obtained recombinant forms of GPC carrying common Gerbich antigens may be useful in serology, and also as model molecules for structure-function studies.     

A,B blood group antigens in tissues of AB heterozygotes. Emphasis on normal and neoplastic urothelium.

The tissue distribution of the A and B blood group antigens was studied in 41 individuals with the heterozygous AB red blood cell (RBC) phenotype. A total of 134 biopsies from a variety of normal tissues (94 from urothelium and 40 from other tissues) were examined. In addition, changes in the expression of these antigens associated with neoplastic transformation were evaluated in 70 biopsies from transitional cell carcinomas of 19 AB heterozygous patients. There was heterogeneity in the distribution of tissue A and B antigens, depending on the cell type, as well as among cells of the same type. Ninety-one percent of AB heterozygotes expressed both A and B antigens in normal epithelial cells, with a mosaic distribution clearly apparent in 50% of these individuals. In 21% of these subjects, the A antigen was undetectable in the vascular endothelial cells in all biopsies from several organs. In most (79%) transitional cell neoplasms, only one of the two antigens was consistently expressed. The results of this study may have implications for the clonal or specific gene deletion theories of neoplasia. They also demonstrate the existence of a subgroup of AB individuals in whom the A antigen is absent specifically from the vascular endothelium.     

Expression of blood group antigens in urinary tract tumours: prospective fluorescence study using cryostat sections of fresh frozen tissues.

Cryostat sections of fresh frozen tissues were used in a prospective study of blood group H and A antigen fluorescence in 73 transitional cell carcinomas of the bladder. The aim was to evaluate antigen expression without subjecting the tumour tissues to organic solvents that extract blood group active glycolipids. Deletion of the genetically predicted antigen was twice as common in tumours of pT1 or greater stage than those of pTa stage and also twice as common in poorly differentiated than in moderately well differentiated tumours. The considerable heterogeneity and overlap, however, in patterns of reactivity in tumours of various histopathological stages and grades and the effect of secretor status on antigenicity meant that there was no obvious antigenic feature that correlated precisely with invasive stage or differentiation grade. It remains to be determined whether the antigen positive and antigen negative tumours represent different disease entities with differing clinical courses. Our results indicate, however, that studies of the blood group antigens in urinary tract tumours are more likely to be of value in research into biochemical disorders in the neoplastic process than in routine clinical assessment as a guide to treatment.     

Acquisition of human blood group antigens by Schistosoma mansoni.

Juvenile forms of Schistosoma mansoni (schistosomula) have been cultured in human blood of various specificities and tested for the presence of blood group substances on their surfaces. The tests employed were survival following transfer into rhesus monkeys immunized against human blood substances, mixed agglutination reactions, and immunofluorescence. A, B, H AND Lewisb+ antigens were expressed at the surface when the parasites were cultured in blood of appropriate specificities. Rhesus, M N S, AND Duffy antigens could not be detected on the parasite surface following culture. The evidence suggests that the expressed blood group antigens are of host origin and are acquired by the parasite during culture, probably in the form of glycolipids or megaloglycolipids. It is likely that these substances are also acquired by parasites in the bloodstream of man. They may serve to mask surface parasite antigens, and so enable schistosomes to evade parasite-specific humoral or cellular immune responses.     

Altered expression of Lewis blood group and related antigens in fetal,normal adult and malignant tissues of the uterine endometrium

Summary The expression of the Lewis blood group and its related antigens in fetal, normal adult and malignant tissues of the uterine endometrium was examined immunohistochemically using a panel of mouse monoclonal antibodies with specificities for Lewis-a (La), Sialyl Lewis-a (SLa), Lewis-b (Lb), Lewis-X (LX), Sialyl Lewis-X (SLX) and Lewis-Y (LY) antigens. La, SLa and SLX having one fucose residue were detected in a small number of fetal tissues, while Lb and LY having two fucose residues were found in most cases. In the adult endometrium, expression of Lb and LY was considerably lower than those in fetal tissues, although expression of La and SLa was not different between these two tissues. Expression of LX and SLX was pronouned in adult when compared with fetal tissues. Malignant endometrial glands expressed La, SLa, Lb and LY, extensively, while LX and SLX were expressed less than in normal tissues.Lb and LY can thus be considered oncofetal antigens, extensively expressed in fetal and malignant tissues but not in normal adult tissues. Expression of Lb and LY was greater than that of La and SLA in carcinoma; an increase in the activity of fucose transferase might be associated with malignant transformation in the uterine endometrium.Supported in part by a Grant-in-Aid for Scientific Research (62480346) from the Ministry of Health and Welfare     

膜联蛋白A7在人宫颈鳞癌和正常组织中的表达

目的 探讨膜联蛋白A7在人宫颈鳞癌和正常组织中的表达情况. 方法 应用免疫组织化学SP法,检测人宫颈鳞癌和正常宫颈组织蜡块各25例中膜联蛋白A7的表达差异;Western .blotting和半定量RT-PCR法检测人宫颈鳞癌和正常组织新鲜标本各25例中膜联蛋白A7的表达差异. 结果 膜联蛋白A7在组织中蛋白和mRNA水平表达一致,均为宫颈鳞癌组织表达高于正常宫颈组织,差异有统计学意义. 结论 膜联蛋白A7在人宫颈鳞癌表达上调, 可能成为宫颈癌诊断的一个新的指示靶点.     

Structural and functional diversity of blood group antigens.

Biochemical and molecular genetic studies have revealed that blood group antigens are present on cell surface molecules of wide structural diversity, including carbohydrate epitopes on glycoproteins and/or glycolipids, and peptide antigens on proteins inserted within the membrane via single or multi-pass transmembrane domains, or via glycosylphosphatidylinositol linkages. These studies have also shown that some blood group antigens are carried by complexes consisting of several membrane components which may be lacking or severely deficient in rare blood group 'null' phenotypes. In addition, although all blood group antigens are serologically detectable on red blood cells (RBCs), most of them are also expressed in non-erythroid tissues, raising further questions on their physiological function under normal and pathological conditions. In addition to their structural diversity, blood group antigens also possess wide functional diversity, and can be schematically subdivided into five classes: i) transporters and channels; ii) receptors for ligands, viruses, bacteria and parasites; iii) adhesion molecules; iv) enzymes; and v) structural proteins. The purpose of this review is to summarize recent findings on these molecules, and in particular to illustrate the existing structure-function relationships.     

Localization of common (cross-reacting) antigens with Neisseria perflava and Klebsiella pneumoniae in tissues of the human bronchopulmonary apparatus

Antigenic similarity was studied between the microsomal fraction of tissues of the human bronchopulmonary apparatus and bacterial cells living in the respiratory tract:Neisseria perflava andKlebsiella pneumoniae. Cross reactions were studied with antimicrosomal sera in the complement fixation test withN. perflava andK. pneumoniae. Fixation of antibacterial antibodies and antibodies against microsomal fractions of the lung tissues was investigated in tissue sections of the human lungs and bronchi. The presence of antigens cross reacting with the antimicrobial sera was demonstrated in the microsomal fraction of tissues of the human bronchopulmonary apparatus.Allergologic Research Laboratory, Academy of Medical Sciences of the USSR, Moscow. Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 81, No. 2, pp. 210–212, February, 1976.     

Expression of blood group-related carbohydrate antigens in normal human pancreatic tissue

The expression of type 1, 2 and 3 chain carbohydrate structures in 15 normal pancreata was investigated by immunohistochemical methods using well-defined monoclonal antibodies. Surgical biopsies of pancreata were obtained from kidney donors while the organs were still perfused. Type 1 chain structures were abundantly expressed including monosialylated(ms)- and disialylated(ds)-Le(a) antigens, which have previously been associated with cancer. Type 2 chain structures were represented by H and Le(y) antigens and to a lesser extent by the precursor structure N-acetyllactosamine, whereas Le(x), dimeric Le(x), and ms-Le(x) were only sporadically observed, in contrast to fetal pancreatic tissue, in which Le(x) has been found to be abundantly expressed. H chain 3 antigen was found in nearly all specimens, whereas precursor structures Tn, sial-Tn and T antigens were absent. Desialylation unmasked the T antigen in all specimens. Absence/masking of Tn, T and related antigens is of special interest, since these antigens are associated with tumor development in other tissues, and may be of importance in pancreatic cancer diagnostics in the future.     

Expression of blood group antigens and lectin binding in adenocarcinomas of the endometrium

To investigate cytoplasmic alterations in cancers of the endometrium, monoclonal antibodies against H, A and B blood group substances and related lectins such as Ulex europaeus agglutinin 1 (UEA-1), Lotus tetragonolobus agglutinin (LTA), and Griffonia simplicifolia 1 (GS1-A4, -B4) were applied to 42 cases of endometrial adenocarcinoma (EAC) and 11 adenomyoses. The control specimens consisted of normal endometria with leiomyoma or adenomyosis from patients of known blood group type. The brightest and most consistent staining was obtained with GS1-A4, showing 98% positivity in EAC in contrast to 18% in normal endometria. In contrast, expression of A antigen was seen in only 24% of EAC and in 13% of normal endometria, UEA-1 binding was seen in 81% of EAC and in 10% of normal endometria, whereas H antigen was seen in 60% of EAC and 0% of normal endometria. In adenocarcinomas, staining was seen not only along the luminal border of glands but also in the cytoplasm and the lateral or basal cell membranes, whereas in the normal endometrium staining was mostly along the luminal border. Thus, a difference in the positivity and localization of glycoconjugates was observed in neoplastic and non-neoplastic endometrium.     

Expression of native and deglycosylated colon cancer mucin antigens in normal and malignant epithelial tissues

Immunohistochemical techniques were used to determine the distribution and cellular location of the mature and precursor forms of a colonic-type mucin in normal and malignant epithelial tissues. The antisera used in this study were prepared against native human colon cancer mucin (LS), partially deglycosylated mucin (HFA or GalNAc-apomucin), and fully deglycosylated mucin (HFB or apomucin). These antisera reacted with most mucin-producing cells of the normal gastrointestinal tract, salivary ductular cells, bronchial epithelial cells, some bronchial mucous glands, and squamous epithelial cells of the esophagus. Breast, endometrium, ovary, prostate, liver, and thyroid were nonreactive. In most normal organs, HFB reactivity was present in the supranuclear and perinuclear cytoplasm and LS and HFA were located primarily in goblet cell vacuoles, apical cytoplasm, and luminal secretions. These findings are consistent with the expected subcellular locations of apomucin and more "mature" mucins. LS, HFA, and HFB were frequently expressed in adenocarcinomas of the colon, stomach, pancreas, and lung. Lymphoma, sarcoma, and melanoma specimens were nonreactive. Alterations in the expression of these mucin antigens in malignant tissues included loss of subcellular compartmentalization, increased intensity of staining, and disappearance of staining. In addition, de novo expression of HFB was observed in one of five breast carcinomas and three of five ovarian mucinous cystadenocarcinomas. These data demonstrate that LS, HFA, and HFB are useful for studying the organ specificities and biosynthetic pathways of one type of mucin in normal and malignant tissues.     

Alloimmunization by blood group antigens from bone allografts

The purpose of this report is to heighten awareness of the risk of blood group antigen sensitization following hone allografting. Two Rh-negative females of childbearing age developed multiple antibodies to Rh antigens following transplantation of bone from Rh-positive donors. A previous pregnancy and/or blood transfusions were ruled out as factors influencing the antibody production. It is postulated that red cells or red cell stroma in the allografts stimulated the antibody production. Therefore, young, Rh-negative bone allograft recipients at risk (especially women) should receive Rh-negative bone allografts. Alternatively, they should receive Rh immune globulin at the time of transplanting if the allograft is from an Rh-positive donor. Bone allografts processed to deplete the hemopoietic marrow and red cells should minimize the risk of blood group antibody sensitization.     

Expression of blood group antigens including HLA markers in human adult liver

The localisation of the principal blood group antigens has been studied in human liver. These blood group antigens included the erythrocyte antigens and the antigen of the major histocompatibility complex. This study was performed by the indirect immunofluorescence technique using polyclonal antibodies of human or animal origin and monoclonal antibodies from hybridomas. This study has shown that the normal hepatocyte is lacking in blood group antigens. On the contrary, the biliary cell was rich in antigenic markers: the main antigens expressed were Lewis, Pr, HLA-A and B antigens. In Kupffer cells, only i and HLA-DR antigens were clearly expressed. The endothelial cells of blood vessels mainly show A, B, H, HLA-A and B antigens; HLA-DR and Pr are slightly expressed. HLA-DR antigens were more strongly expressed on veins than on arteries. Dendritic cells have been identified in the portal space of human liver. They bore i and HLA-DR antigens.     

Localization of common (cross-reacting) antigens with tissues of the human bronchopulmonary apparatus in cells ofNeisseria perflava andKlebsiella pneumoniae

The localization of common antigens with tissues of the human bronchopulmonary apparatus was studied in cells ofNeisseria perflava andKlebsiella pneumoniae. Cross reactions of several structures ofN. perflava andK. pneumoniae cells (capsule, cell walls, fractions of cytoplasmic structures, hyaloplasm) were studied in the complement fixation test (CFT) with antilung sera. Antigens cross-reacting with antilung sera were found not only in surface structures (cell walls) of the bacterial cells but also in deep components (cytoplasmic fraction rich in RNP) of the microorganism.Allergologic Research Laboratory, Academy of Medical Sciences of the USSR, Moscow. Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 81, No. 3, pp. 349–350, March, 1976.     

Trachea-specific antigens in normal and malignant human tissues

One to four trachea-specific antigens were demonstrated regularly by means of various rabbit antisera to human tracheal mucosa, using immunoelectrophoresis or Ouchterlony tests. In addition, one or more antigens common to trachea and oesophagus were found. Trachea-specific antisera cross-reacted with rhesus monkey but not with bovine, ovine, equine or porcine trachea. The three most prominent trachea-specific antigens were localized in the same fraction by chromatography. The chromatographic fraction was usable in tanned cell haemagglutination, but only trachea-oesophagus antigen, and not trachea-specific antigen, seemed to attach to tanned cells. Of the ten cancers of the respiratory tract tested, three were found to retain some trachea-specific antigen.     

Universal red blood cells--enzymatic conversion of blood group A and B antigens.

Accidental transfusion of ABO-incompatible red blood cells (RBCs) is a leading cause of fatal transfusion reactions. To prevent this and to create a universal blood supply, the idea of converting blood group A and B antigens to H using specific exo-glycosidases capable of removing the immunodominant sugar residues was pioneered by Goldstein and colleagues at the New York Blood Center in the early 1980s. Conversion of group B RBCs to O was initially carried out with alpha-galactosidase extracted from coffee beans. These enzyme-converted O (ECO) RBCs appeared to survive normally in all recipients independent of blood group. The clinical trials moved from small infusions to single RBC units and finally multiple and repeated transfusions. A successful phase II trial utilizing recombinant enzyme was reported by Kruskall and colleagues in 2000. Enzymatic conversion of group A RBCs has lagged behind due to lack of appropriate glycosidases and the more complex nature of A antigens. Identification of novel bacterial glycosidases with improved kinetic properties and specificities for the A and B antigens has greatly advanced the field. Conversion of group A RBCs can be achieved with improved glycosidases and the conversion conditions for both A and B antigens optimized to use more cost-efficient quantities of enzymes and gentler conditions including neutral pH and short incubation times at room temperature. Of the different strategies envisioned to create a universal blood supply, the ECO concept is the only one, for which human clinical trials have been performed. This paper discusses some biochemical and clinical aspects of this developing technology.