Immunolocalization of cell-wall-deficient forms of Mycobacterium tuberculosis complex in sarcoidosis and in sinus histiocytosis of lymph nodes draining carcinoma.

In sarcoidosis, pleomorphic chromogens (PCs) occur as multivariate pigmented elements within sinusoids of lymph nodes (sinusoidal phase) and as tiny "round bodies" detectable in granulomas (generalized phase). The sinusoidal phase occurs in other conditions as well and characteristically contains yeastlike bodies also known as H-W bodies. To elucidate the antigenic profile of all variant forms, 28 cases of sarcoidosis (series A) and 14 cases of malignancy associated sinus histiocytosis (series B) were studied immunohistochemically with panels of various antibodies, including antimycobacterial MAbs specific for M tuberculosis complex (TB68, TB71), for M. leprae (MMP-I-3C3) and for cross-reactive mycobacterial antigens (F24-2-3 and F116-5, the latter recognizing superoxide dismutase). Results for series A indicate that: 1) PCs are cell-wall-deficient (CWD) mycobacterial forms belonging to M. tuberculosis complex (over 95%); 2) both phases are antigenically identical parts of the L-cycle; 3) "round bodies" of the "infective" phase have an endolysosomal evolution; 4)uncommon vacuolated forms represent a labile spheroplast stage; 5) the yeastlike bodies are specialized sinusoidal large bodies of unknown function. Results for series B show that in roughly two thirds of cases the pigmented forms are also CWD mycobacteria, have the same immunophenotype as sarcoid PCs in 35.7% of cases, have a much higher incidence of labile vacuolated forms and, finally, that malignancy associated "pseudosarcoid" granulomas do not differ antigenically from genuine sarcoid granulomas. Unlike conventional mycobacteria, PCs do not express cytoskeletal proteins consistently. Their general reactivity for HBcAg raises the possibility of phage interactions being responsible for the L-cycle since it may reflect shared epitopes between unrelated virus entities.

Cross-reactive and species specific Mycobacterium tuberculosis antigens in the immunoprofile of Schaumann bodies: a major clue to the etiology of sarcoidosis.

Sarcoidosis, once thought to be a variant of tuberculosis, is currently listed as a disease of unknown etiology. The present study was initiated by unpublished observations that Schaumann bodies-the laminated inclusions often encountered in sarcoid granulomas-cross-reacted with commercial polyclonal antibodies to Mycobacterium bovis, Mycobacterium duvalii and Mycobacterium paratuberculosis. Given the broad cross-reactivity of many mycobacterial antigens, those findings lacked specificity but warranted in depth probing of the immunoprofile of the bodies, particularly for specific mycobacterial antigens. Formalin-fixed tissue from eight patients with an established diagnosis of sarcoidosis was studied with panels of antibodies against both common cytoplasmic proteins and various mycobacterial antigens, using a labeled streptavidin-biotin-alkaline phosphatase technique. Our findings indicate that Schaumann bodies are indeed residual bodies of heterophagic mycobacterial derivation. They immunostained intensely for the lysosomal proteins muramidase and CD68, variably for some cytoskeletal proteins (tubulin, desmin, vimentin) and not at all for cytokeratin, muscle actin, alpha-1-antichymotrypsin and ferritin. Both cross-reactive and species specific antigenic determinants of M. tuberculosis complex were shown to be present. Affinity absorption with killed intact bacilli H37 Rv resulted in virtually equal loss of binding by all polyclonal antimycobacterial antibodies to cross-reactive ligands in Schaumann bodies. In addition, the bodies were clearly labeled with the monoclonal antibodies TB68 and TB71, known to recognize species specific epitopes of Mycobacterium tuberculosis complex. Although obtained on a small number of cases, our findings uphold Schaumann's original postulate that the laminated calcific inclusions represent remnants of "transformed tubercle bacilli".

Asteroid bodies: products of unusual microtubule dynamics in monocyte-derived giant cells. An immunohistochemical study.

We have studied asteroid bodies (ABs) of multinucleated giant cells (MGCs) in a series of sarcoid and foreign body granulomas with a standard streptavidin-biotin peroxidase technique, using commercial antibodies against collagen, vimentin and tubulin on routinely processed tissue as well as, in one case, on fresh frozen sections (FS). Our findings clearly indicate that ABs are products of the microtubule (MT) system and lack collagen. The tubulin in them stains in fresh FS but is "masked" in formalin-fixed tissue. It can be fully "unmasked" by dephosphorylation and partially by trypsinization. Compared to single microtubule organizing centers (MTOCs) in mononuclear cells serving as internal controls, ABs are obvious replicas of centrosome-nucleated MT assemblies from which they differ principally by the disproportionate size of their components and by the invariable vacuolation of the surrounding cytoplasm. Relying on bits of relevant information gleaned from the literature, our observations support the following preliminary conclusions: 1) spokes are massive bundles of MTs rich in tyrosinated alpha-tubulin coassembled in phosphorylated linkages with yet unidentified microtubule associated proteins (MAPs) and probably microfilament proteins; cores are masses of pericentriolar material including amorphous tubulins, MAPs, phosphoproteins and phospholipids; 2) their size, at least in some ABs, appears to indicate the presence of overlapping centrosome-nucleated MTOCs which in monocyte-derived MGCs are known to be multiple; 3) the cytoplasmic vacuolations around them reflect a collapse and retraction of intermediate filaments (IFs), indicating substantial ongoing MT depolymerization with disruption of MT-IF interactions; 4) ABs are products of unusual MTOC dynamics characterized by simultaneous MT assembly and depolymerization; such a phenomenon, termed "microtubule catastrophe", has been recognized in vitro with centrosome-nucleated MT assemblies under conditions of low tubulin concentrations.

Selective immunolabeling of early gestational cytotrophoblast and its neoplastic counterpart by the monoclonal antibody Ber-EP4.

This report presents preliminary observations on the distribution of an epithelial surface antigen in human trophoblastic cells studied with the monoclonal antibody Ber-EP4 by means of an immunoperoxidase technique. Three cases of gestational choriocarcinoma and 20 cases of both villous and extravillous trophoblast, including one complete hydatidiform mole and one exaggerated placental site, were examined with a panel of monoclonal antibodies against epithelial antigen (Ber-EP4), cytokeratin (AE1 and AE3), vimentin (VIM), carcinoembryonic antigen (CEA) and epithelial membrane antigen (EMA), as well as with polyclonal antibodies against the beta subunit of human chorionic gonadotropin (hCG), human placental lactogen (hPL) and placental alkaline phosphatase (P1AP). While, as anticipated, all currently defined trophoblastic cell lines expressed low molecular weight cytokeratin only first trimester cytotrophoblastic cells and their neoplastic counterpart were found to possess the epithelial antigen detected by Ber-EP4. This distinctive property of the cytotrophoblast declined rapidly after eight weeks of gestational age but persisted in second trimester molar tissue and was prominently displayed in choriocarcinoma. By identifying the presence of cytotrophoblastic cell lines in malignancy more reliably than can be achieved with conventional panels of hCG, hPL and P1AP, Ber-EP4 has potential diagnostic utility in gestational trophoblastic disease. Our findings also suggest that the monoclonal anti-CEA antibody may be helpful in distinguishing between syncytiotrophoblastic cells and non-reactive multinucleate cells of the intermediate trophoblast.

Mucinous neoplasm in the cervix associated with a mucinous neoplasm in the ovary and concurrent bilateral sex cord tumors with annular tubules: immunohistochemical study.

The patient described synchronous mucinous tumors of the cervix and ovary and concurrent annular tubules, but without the classical stigmata of Peutz-Jeghers syndrome. The cervical tumor was an invasive mucinous adenocarcinoma with mixed components of minimal deviation and less-well-differentiated endometrioid morphology. The ovarian tumor had the benign appearance of a mucinous adenoma but histologically revealed areas of invasive carcinoma. Immunohistochemical studies of the mucinous neoplasms of the cervix and ovary are discussed. Neither the staining properties of mucin, the pattern of immunostaining for carcinoembryonic antigen, nor any other common markers were helpful in distinguishing the mucinous neoplasms. Positive immunostaining for low-molecular-weight cytokeratin in the filament profile of sex cord tumors with annular tubules was of particular interest since it has not to our knowledge been previously described.

Carcinoid tumors.

The authors carried out a retrospective study of 32 patients (23 M, 9 F) with carcinoid tumors who were diagnosed and treated at Harlem Hospital Center, New York, from 1967 to 1988. All the patients were black and the commonest sites were the ileum (28.1%), rectosigmoid and rectum (21.9%), and the appendix and lung (15.6% each). Metastasis correlated with site, size, and depth of the primary tumor and occurred in 12 patients (38%), most frequently to the regional lymph nodes and liver. Carcinoid syndrome developed in 12.5% (3 F, 1 M). Surgical resection for cure or palliation was the mainstay of treatment. Overall 5 year survival rate was 66%, and for those with metastases was 0%. The poorer survival rates are probably related to the socioeconomic status of our patient population. The only observed racial difference compared to other series is the preponderance of males, and the disproportionately higher ratio of females with the carcinoid syndrome.

Histological and ultrastructural findings in a case of the Sézary syndrome.

This paper deals with the histological and ultrastructural findings in a case of the Sézary syndrome. The striking nuclear and cytoplasmic features of the Sézary cell are illustrated, and the similarities of this cell to the mycosis fungoides cell are once more stressed.