The IgM antibody level against ganglioside GM2 correlates to the disease status of HIV-1-infected patients

HIV-1 infection induces the expression of high level of GM2 ganglioside on infected cells and IgM antibody (Ab) against GM2 can cause complement (C)-mediated cytolysis of HIV-1-infected cells. Since GM2 is immunogenic in human, we proposed that an anti-GM2 IgM Ab may be produced by some HIV-1-infected patients and the titer of this Ab might provide some insight into the progress of the disease. On this premise, the amount of IgM Ab against GM2 was determined in 124 HIV-1-infected patients and 111 seronegative donors. As expected, the anti-GM2 IgM Ab titers of the patients was significantly higher than that of the seronegative donors while the total IgM levels remained unchanged. In addition, we determined the CD4+ cell count and the HIV-RNA load in the HIV-1-infected patients. The results showed a positive correlation between the anti-GM2 IgM Ab titer and CD4+ cell count but a negative correlation between the anti-GM2 IgM Ab titer and HIV-RNA load. These suggest that anti-GM2 IgM Ab induced and/or enhanced by HIV-1 infection causes C-mediated cytolysis of HIV-1-infected cells in vivo to a certain extent, and may help lower the plateau level of the HIV-RNA load. Therefore, the amount of IgM Ab against GM2 may be related to the prognosis of HIV-1 infected patients.     

Selective gene therapy for prostate cancer cells using liposomes conjugated with IgM type monoclonal antibody against prostate-specific membrane antigen

Prostate cancer cells express prostate-specific membrane antigen (PSMA). We developed an IgM type monoclonal antibody against PSMA. The antibody was coupled to poly-L-lysine and thereafter this conjugate was mixed with cationic liposomes containing plasmid DNA. The antibody-liposome complex was tested whether it could deliver the gene of interest selectively to the PSMA positive cells. As assessed by beta-galactosidase reporter gene, the transfection efficiency was 13.2% with anti-PSMA-liposome complex as compared to 4% with control IgM liposome complex. In contrast, no such differences were observed in PSMA negative PC-3, DU145 and T24 cells. Furthermore, in the suicide gene therapy in vitro with thymidine kinase gene plus ganciclovir system, anti-PSMA liposome complex demonstrated a selective growth inhibitory effect on PSMA positive LNCaP cells but not on PSMA negative cell lines.     

O-acetyl sialic acid specific IgM in childhood acute lymphoblastic leukaemia

Initial studies have revealed an enhanced surface expression of O-acetylated sialoglycoconjugates (O-AcSGs) on lymphoblasts concomitant with high titres of IgG in childhood Acute Lymphoblastic Leukaemia (ALL) (Mandal C, Chatterjee M, Sinha D, Br J Haematol 110, 801–12, 2000). In our efforts to identify disease specific markers for ALL, we have affinity-purified IgM directed against O-AcSGs that reacts with three disease specific O-AcSGs present on membrane proteins derived from peripheral blood mononuclear cells (PBMC) of ALL patients. Antibody specificity towards O-AcSGs was confirmed by selective binding to erythrocytes bearing surface O-AcSGs, decreased binding with de-O-acetylated BSM and following pretreatment with O-acetyl esterase. Competitive inhibition ELISA demonstrated a higher avidity of IgM for O-AcSG than IgG. Flow cytometry demonstrated the diagnostic potential of purified O-AcSA IgM as binding was specific with ALL patients and minimal with other haematological disorders and normal individuals. It therefore may be adopted as a non-invasive approach for detection of childhood ALL. Taken together, the data indicates that carbohydrate epitopes having terminal O-AcSA 2 6 GalNAc determinants induce disease specific IgG and IgM, potentially useful molecular markers for childhood ALL.     

Binding of synthetic peptides by a human monoclonal IgM with an unusual combining site structure

Using X-ray crystallography, a human monoclonal IgM cryoglobulin (Mez) was found to have an unusual combining site topography. Analysis of the unliganded Fv at 2.6 A resolution revealed that the HCDR3 had partitioned the active site into two compartments [Ramsland PA et al. 2000. Mol. Immunol. 37: 295-310]. The two cavities had dimensions and chemical properties that were compatible with the binding of peptides. In this study, libraries of peptides were prepared using solid-phase synthesis. Binding of the intact Mez IgM to these peptides was tested by enzyme-linked immunoassays. Screening of 400 dipeptides revealed that binding was markedly skewed toward amino acids with aromatic side-chains (Phe and Trp), especially when located in the second position. Preferential recognition of aromatic side-chains by Mez IgM was confirmed with larger peptides of three to five residues, but C-terminal positioning was not favored in these peptides. Mez IgM also showed binding propensities for acidic residues (Asp and Glu) as well as several other side-chains with different chemical properties, including His, Pro, Asn and Gln. Mez IgM recognized sets of overlapping octapeptides representing the sequences of the constant domains of human IgG1 heavy chains. These peptides represented similar stretches of polypeptide on the three-dimensional structures of all three constant domains (CH1, CH2 and CH3). Thus, Mez IgM may recognize structurally homologous regions of immunoglobulin domains, which were conserved during the evolution of the immune system.     

Natural antibodies as a sensor of electronegative damage-associated molecular patterns (DAMPs)

Natural antibodies (Abs), predominantly IgMs, play an important function in the host response to the recognition of endogenous danger signals called damage-associated molecular patterns (DAMPs). Many of the natural IgM Abs also show several different antigenic cross-reactivities toward covalently modified proteins, such as oxidized low-density lipoproteins and advanced glycation end products. Of interest, a recent study has shown that these DAMPs have several physicochemical characteristics that differ from native proteins, such as an increased negative charge due to modification of the lysine residues. This finding may provide a mechanistic insight into the multispecificity of the natural Abs.     

The influence of 2-mercaptoethanol (2-ME) treatment of IgG antibody on its ability to induce cytotoxicity in nonsensitized lymphocytes

Administering dextran 2 hr before giving antigen (sheep red blood cells) to X-irradiated mice repopulated with B and T cells caused alterations in antibody synthesis. Besides a slight increase in the number of cells making IgM antibodies, cells producing IgG antibodies were detected at a time when none are usually present in untreated control animals.Repopulating X-irradiated mice with B cells treated with dextran either in vivo or in vitro and immunizing with sheep red blood cells resulted in twice the background number of sheep red blood cell-specific IgM-plaque-forming cells at 4.5 days of immunization as well as a small number of IgG-plaque-forming cells at 8.5 days. At these times, control animals given only B cells and sheep red blood cells possessed a background number of IgM-plaque forming cells and no IgG plaque-forming cells.Incubating B cells with θ-specific antiserum and complement to remove residual T cells prior to transplantation obliterated dextran's stimulus. Dextran's alterations of immunological responses towards unrelated antigens therefore appears to be manifested through T cells. The latter must be in company with B cells at the time of exposure to dextran. Thus, T cells upon contact with dextran apparently release B cell stimulatory factor (s) responsible for increasing the number of IgM-forming cells and for triggering IgG-forming cells.     

Proteostasis and “redoxtasis” in the secretory pathway: Tales of tails from ERp44 and immunoglobulins

In multicellular organisms, some cells are given the task of secreting huge quantities of proteins. To comply with their duty, they generally equip themselves with a highly developed endoplasmic reticulum (ER) and downstream organelles in the secretory pathway. These professional secretors face paramount proteostatic challenges in that they need to couple efficiency and fidelity in their secretory processes. On one hand, stringent quality control (QC) mechanisms operate from the ER onward to check the integrity of the secretome. On the other, the pressure to secrete can be overwhelming, as for instance on antibody-producing cells during infection. Maintaining homeostasis is particularly hard when the products to be released contain disulfide bonds, because oxidative folding entails production of reactive oxygen species. How are redox homeostasis (“redoxtasis”) and proteostasis maintained despite the massive fluxes of cargo proteins traversing the pathway? Here we describe recent findings on how ERp44, a multifunctional chaperone of the secretory pathway, can modulate these processes integrating protein QC, redoxtasis, and calcium signaling.     

九项呼吸道感染病原体IgM检测结果分析

目的 探讨新乡市第一人民医院呼吸系统感染患者9种呼吸道病毒的分布情况.方法 回顾性分析该院门诊和住院的呼吸系统感染者中接受病毒抗体检测330例患者的检测结果.结果 入选病例共330例,检测出抗体阳性者193例,阳性检出率为58.48％,其中肺炎支原体(MP) 131例(39.70％),嗜肺军团菌(LP) 89例(26.97％),肺炎衣原体(CPn) 17例(5.15％),Q热立克次体(COX)15例(4.55％),副流感病毒(PIVs)7例(2.12％),呼吸道合胞病毒(RSV)6例(1.82％),甲型流感病毒(INFA)5例(1.52％),乙型流感病毒(INFB)3例(0.91％),腺病毒(ADV)2例(0.61％).结论 MP、LP、CPn分居呼吸道病毒感染的前三位;除CPn以秋季感染明显高于春冬夏季节;其余病毒感染无明显季节、年龄、性别差异.     

A human IgM signals axon outgrowth: coupling lipid raft to microtubules

Mouse and human IgMs support neurite extension from primary cerebellar granule neurons. In this study using primary hippocampal and cortical neurons, we demonstrate that a recombinant human IgM, rHIgM12, promotes axon outgrowth by coupling membrane domains (lipid rafts) to microtubules. rHIgM12 binds to the surface of neuron and induces clustering of cholesterol and ganglioside GM1. After cell binding and membrane fractionation, rHIgM12 gets segregated into two pools, one associated with lipid raft fractions and the other with the detergent-insoluble cytoskeleton-containing pellet. Membrane-bound rHIgM12 co-localized with microtubules and co-immuno precipitated with β3-tubulin. rHIgM12-membrane interaction also enhanced the tyrosination of α-tubulin indicating a stabilization of new neurites. When presented as a substrate, rHIgM12 induced axon outgrowth from primary neurons. We now demonstrate that a recombinant human mAb can induce signals in neurons that regulate membrane lipids and microtubule dynamics required for axon extension. We propose that the pentameric structure of the IgM is critical to cross-link membrane lipids and proteins resulting in signaling cascades.     

A long term experimental study of canine visceral leishmaniasis

Previous studies on Leishmania infantum and the canine immune response are derived mainly from short-term studies. To date, there have been no longitudinal studies that perform a serial analysis of the intensity of infection in conjunction with immunological parameters and clinical signs in Leishmania-infected dogs. For this purpose, six dogs were infected experimentally by the i.v. route and were monitored for 1 year. Clinical, immunological (humoral and cellular response) and parasitological (parasitaemia) parameters were evaluated monthly. Four dogs developed clinico-pathological signs compatible with leishmaniasis, whereas two dogs showed few abnormalities during the study. Evaluation of clinical, immunological and parasitological parameters showed that the intensity of Leishmania infection in blood samples, as indicated by the amount of Leishmania DNA, was correlated significantly with IgG, IgG1, IgG2, IgA, and IgM concentrations and with clinical signs. Parasitaemia and Leishmania-specific cell-mediated immunity were inversely correlated. Moreover, higher quantities of Leishmania DNA were detected in the liver, spleen, lymph node, skin and bone marrow of dogs exhibiting clinical signs than those exhibiting few such signs. These findings suggest that progressive disease in experimental canine leishmaniasis is associated with specific T-cell unresponsiveness and unprotective humoral responses which allow the dissemination and multiplication of L. infantum in different tissues.