Cytologic and immunologic prognosis of infertility. I. Prognostic significance of cytologic study of peritoneal fluid of infertile patients

Cytological investigation of the peritoneal fluid from 110 patients was performed. The fluid was taken during laparoscopical surgery. Cytological patterns of the peritoneal fluid from women with tube-peritoneal infertility, myoma utery, and ovarian cysts in combination with infertility were described. Cytological data on the peritoneal fluid can serve one of the criteria of the state of reproductive organs and future prognostication in infertal patients.     

Blood levels of immunoglobulins G,A,M, circulating immune complexes, complement and proteins in patients on intermittent peritoneal dialysis

The study involved 17 patients on IPD. Blood serum levels of IgG, IgA, IgM, circulating immune complexes, complement and proteins were determinated at the beginning of therapy, after 3, 6, 12 months on IPD and after 1 year on hemodialysis. The frequency of peritonitis was noted during this time. Peritonitis was the most frequent during first 3 months on IPD. No differences in blood serum levels of IgG, IgA, IgM, in the specific periods of IPD were noted. A significant increase in blood serum circulating immune complexes in patients on IPD and hemodialysis compared to the control group was found. A significant decrease in blood serum of C3 complement in patients on IPD and hemodialysis in comparison with the controls were found. A significant decrease in blood serum proteins at the beginning of IPD and after 3 months IPD in comparison with proteins concentration in patients on hemodialysis was observed.     

Isotype-specific rabbit antibodies against chinchilla immunoglobulins G, M, and A.

Chinchillas have become a preferred animal model for studying otitis media, and are also useful in studying insulin release, gastrin physiology, intestinal infection, and hepatocellular pathophysiology. Immunopathologic studies in the model, however, have been limited by absence of specific antibody reagents against chinchilla immunoglobulins. We describe a method for preparing isotype-specific rabbit antibodies against the heavy-chain components of chinchilla immunoglobulins G, M, and A. Chromatographic techniques were used to isolate chinchilla immunoglobulins from serum and breast milk; heavy-chain fractions were isolated and used as antigens to produce isotype-specific antibodies in New Zealand White rabbits. Enzyme-linked immunosorbent assay of these antisera disclosed anti-light chain cross-reactivity, which was removed by affinity chromatography. The isolation and affinity purification techniques were highly reproducible. The availability of these reagents should greatly enhance the utility of the chinchilla in modeling human disease.     

Bound class M, A and G immunoglobulins in the thymus of myasthenia gravis patients]

Deposits of granular material containing IgM, IgA, and IgG were revealed in the thymus of patients with myasthenia by direct immunofluorescence. Treatment of the thymus sections by unlabeled preparations against individual classes of human immunoglobulins inhibited the reaction of the granular material with the homologous labeled preparations. Disappearance of fluorescence of these deposits was also seen in treatment of the sections with glycine-HCl-buffer, pH 2.8. These data permitted a suggestion that granular material represented immune complexes where IgM, IgA, and IgG served as antibody, and thymus tissue components--as an antigen. The presence of bound immunoglobulin in the thymus indicated that an autoimmune process directed against the tissues of this organ occurred in myasthenia.     

Cytologic and immunocytochemical studies of cerebrospinal fluid in meningeal sarcoidosis. A case report.

Serial studies were done on cerebrospinal fluid (CSF) from a patient with sarcoidosis involving the meninges. Initially when the disease was active, the CSF protein was increased and glucose decreased. The number of cells in the CSF was moderately increased, and many mononuclear cells were present. Cytologic studies of the CSF showed many normal and some atypical lymphocytes. Immunochemical studies showed that most of these lymphocytes were T cells, with T-helper cells predominating over T-suppressor cells by a ratio of 3.92; B-lymphocytes were polyclonal. Subsequent studies of the CSF over the following three and one-half years showed that the protein and glucose content and the cell counts in the fluid did not correlate well with the activity of the disease. The number of atypical lymphocytes seemed to be a more useful marker of disease activity in the patient. Cytologic studies, when interpreted within the context of other CSF and clinical findings, are useful for the assessment of patients with sarcoidosis involving the meninges.     

Immunohistochemical localization of immunoglobulins A, G and M in the mouse female genital tract

The localization of immunoglobulins A, G and M (IgA, IgG, IgM) in the mouse genital tract was studied by immunoperoxidase techniques at oestrus, on the day of mating and at the time of implantation. In the horns and body of the uterus, IgA and IgG were located in plasma cells in the endometrium surrounding uterine glands and in the gland lumina. The numbers of these plasma cells increased markedly between Day 1 and Days 4 and 5 of pregnancy and the ratio of plasma cells containing IgA and IgG was about 3 or 4 to 1 at all stages. Area measurements indicated that the increased number of plasmacytes was not due to an increase in the amount of endometrial, myometrial or glandular tissues. Plasma cells were not detected in the cervix and vaginal fornix at oestrus and Day 1, but a few were present on Day 5. In the oviduct, plasma cells containing IgA and IgG were present only in the preampulla and both immunoglobulins were present in the extracellular space of the lamina propria only in this region. No IgM was detected in any part of the reproductive tract at any of the times studied. Uteri on Day 1 of pregnancy contained bacteria of several kinds, some of which were aggregated and coated with IgA. This suggests that the uterine lumen at this time may contain specific anti-bacterial IgA antibodies. Our observations indicate that the horns and body of the uterus and the preampulla of the oviduct are major sites of a local immune system in the female mouse genital tract.     

Comparative conformational studies on the tryptic digestion fragments of human immunoglobulins M and G

IgM¹ immunoglobulins were cleaved into Fabμ and (Fc)5μ fragments by tryptic digestion. Comparative circular dichroism studies with the corresponding IgG fragments show that the Fab portions of IgG and IgM proteins have very similar CD spectral features, although the same is not true for their Fc fragments. These studies indicate the presence of higher amount of beta-structured regions in Fcμ than in Fcγ. Also, there are considerable differences in their pH-dependent structural transitions as measured by CD spectral changes. The conformational differences between IgG and IgM immunoglobulins are more pronounced in their Fc portions, which carry out class specific biological functions, rather than in Fab portions, which contain antigen combining sites.     

Localization of immunoglobulins G, A and M in glial cells of reactive and neoplastic origin

The localization of immunoglobulins G, A and M in glial cells of neoplastic and reactive origin have been investigated by the use of the PAP (peroxidase-antiperoxidase) method on paraffin embedded tissue previously fixed in calcium formol. It has been found, that some glial cells of astrocyte type showed a very intense staining when oligoclonal antibodies to human immunoglobulins G, A and M specific for gamma, alpha, and mu chains were used. The localization of immunoglobulins was disclosed in astrocytes of various morphology; astrocytes with well developed processes, gemistocyte type cells without or only with short and thick cell processes and in small cells with scanty cytoplasm. The number of cells with immunoglobulins localized is very small. No positive results have been noted if the normal brain tissue is concerned. The specificity of the method is discussed.     

Prognostic value of cytologic examination of peritoneal washings in patients with endometrial carcinoma

Peritoneal pelvic washings from 54 women with pathologic stage I endometrial carcinoma were evaluated in a blind retrospective fashion for the concentration of malignant cells present. None of the 42 patients with normal washings developed recurrence after a median disease-free survival of 36 months. Of the 12 patients with adenocarcinoma in the washings, 4 had high concentrations of malignant cells (greater than 1000 cells/100 ml sample), and all 4 died as a consequence of carcinoma within two years. The remaining eight patients had lower concentrations of malignant cells in the washings (less than 1000 cells/100 ml sample), and six of these patients had no evidence of disease after 37 to 64 months. Cox's nonparametric statistical model showed that increasing concentrations of adenocarcinoma cells in washings significantly shortened the time to recurrence of disease. The abundance of malignant cells has prognostic importance in identifying those patients with pathologic stage I disease who may require more aggressive therapy.     

Synthesis of immunopoietins--new biologically active low molecular weight fragments of immunoglobulins G, M, E and A

Low-molecular fragments of immunoglobulins IgG-(245-349) (Glu-Pro-Gln-Val-Tyr), IgM-(451-455) (Arg-Pro-Asp-Val-Tyr), IgA-(347-351) (Arg-Pro-Glu-Val-His), and IgE-(430-435) (Ala-Ala-Pro-Glu-Val-Tyr), potentially active immunoregulators of a novel type, have been synthesised by classical methods of peptide chemistry. This group of compounds, which in biological effects are similar to thymopoietin, was given the name of immunopoietins.     

Preparation of immunoglobulins G,A,M (IgGAM) for therapeutic use. Conditions for enrichment in IgA or in IgM]

Antibodies directed against viruses and bacteria are not equally distributed among the main classes of immunoglobulins, e.g. IgG, IgA and IgM. It has been found that IgM is mostly concerned with certain antibacterial activities (Salmonella, Escherichia coli and Pseudomonas) and IgA with high antibody titers for poliomyelitis virus I whereas antibody activities against many viruses such as influenza and measles virus occur preferentially in the IgG population. Furthermore, isolated immunoglobulin deficiency syndromes are actually well known. In the light of these findings, new concepts of immunotherapy have developed. Massive i.v. IgG-therapy is already widely used in congenital and acquired severe hypogammaglobulinemia. Preparations enriched in IgA and IgM are needed to complete the immunotherapeutical possibilities. Such a fraction called IgGAM has already been prepared in our Institute. Fraction III obtained during large scale fractionation is used as starting material and caprylic acid for the precipitation of most proteins other than the immunoglobulins present in fraction III. The immunoglobulin concentrate is finally obtained by ethanol precipitation of the caprylic acid supernatant. The present study is concerned with various modifications of the initial technique in order to obtain fractions more specially enriched in IgA or in IgM. In some cases the standard IgGAM fraction has been submitted to a further fractionation step, such as adsorption of IgG on DEAE-cellulose or precipitation of certain immunoglobulins achieved by Rivanol or by lowering the salt concentration. In other trials the fractionation procedure starting from fraction III has been modified. Rivanol has been used as a precipitating agent for the subfractionation of fraction III. It is well known that IgG is soluble in the presence of Rivanol. This technique was thus used in order to obtain preparations enriched mainly in IgM and IgA. The precipitate obtained after the addition of Rivanol was dissociated by NaCl and the solution further subfractionated by caprylic acid. In a similar way PEG was associated with the caprylic acid precipitation step. PEG precipitates proteins mainly in function of their molecular weight. However, the enrichment of IgM of the final fraction did not exceed 32% and much IgM was lost under the experimental conditions. It proved easiest to suspend fraction III in distilled water leaving IgM in the precipitate; it is dissolved and the solution submitted to a slightly modified caprylic acid precipitation step. This fraction contains 35-40% IgM, few (2-6%) IgA and about 50% IgG whereas an IgA (35%) enriched fraction is obtained when fraction III is solubilized with acetate at pH 6.2 and then submitted to precipitation by caprylic acid under slightly modified conditions as compared with our standard IgGAM. Thus, simple modifications of the standard procedure allow to prepare fractions enriched more specially in IgM or IgA. Fractions poor or almost devoid of IgG can also be obtained...