Anti-measles Antibodies Research Articles

Intraventricular interferon in a case of subacute sclerosing panencephalitis

A patient with subacute sclerosing panencephalitis (SSPE) was treated by direct administration of Interferon B into the cerebral ventricles. Immediately after the first dose intrathecal synthesis of IgG and the CNS antimeasles antibody titer fell significantly, only to rise again to the pretreatment levels 2 months after discontinuation of treatment. There was no improvement in the patient's clinical status.

Isolation of measles virus polypeptides from infected brain tissue by affinity chromatography

A method has been developed to isolate measles virus proteins from infected hamster brain tissue. Suckling hamsters inoculated intracerebrally with the HBS strain of measles virus were used in these studies. Viral proteins were isolated from infected brain lysates by affinity chromatography on Sepharose beads coupled with IgG from rabbit hyperimmune anti-measles serum. The eluted proteins were separated by SDS-polyacrylamide gel electrophoresis (SDS-PAGE), electrophoretically transferred onto blotting matrix, and immunolabelled with anti-measles antibodies. Individual viral proteins were identified by labelling with monoclonal or monospecific antibodies. All viral proteins except the fusion (F 1) protein were identifiable on the immunoblots in relative amounts comparabe to purified virions. In addition, a second phosphoprotein (P) band not found in purified virions was present in infected brains and cell cultures infected with HBS or LEC strains of virus. This method should be useful for isolating small quantities of viral proteins from large amounts of tissue, and should make possible the characterization of measles virus proteins in persistently infected CNS tissue.

Protective Effect of Measles Virus Inoculation on Subacute Sclerosing Panencephalitis Virus‐Infected Mice

The Biken strain of subacute sclerosing panencephalitis (SSPE) virus caused a fatal neurologic disease in adult mice after intracerebral inoculation. However, the mice were completely protected from the disease when a high dose of measles virus was given intracerebrally after the SSPE virus infection. The measles virus inoculation induced interferon production and immune responses. An experiment with athymic nude mice showed that interferon and anti-measles antibody were able to prolong the incubation period of the disease but not to protect the SSPE virus-infected nude mice from death. For complete protection, T lymphocytes appeared to be essential. The present study suggested that the protective effect of measles virus inoculation is basically due to the induction of immune responses and that SSPE virus infection in mice is susceptible to immune reactions.

Antigen mimicry involving measles virus hemagglutinin and human respiratory syncytial virus nucleoprotein

Intergenic antigenic relationships between measles virus and respiratory syncytial (RS) virus-specific structural components were studied by using monoclonal antibodies. Of 75 monoclonal antibodies against these components, only one, an anti-measles virus hemagglutinin monoclonal antibody, cross-reacted. Immunofluorescence analysis of measles virus- and RS virus-infected cells with this monoclonal antibody showed qualitatively different staining patterns which indicated that the antigen involved in cross-reaction was the RS virus nucleoprotein or phosphoprotein. A radioimmunoprecipitation assay showed the antigen to be the nucleoprotein.

A simple method to detect intrathecal production of specific antimeasles antibodies in cerebrospinal fluid during subacute sclerosing panencephalitis

The intrathecal production of antimeasles antibodies was studied using the enzyme-linked immunosorbent assay in eight specimens of serum and cerebrospinal fluid (CSF) from patients with clinical signs of subacute sclerosing panencephalitis (SSPE). The test was performed using a 1:5 dilution of CSF and a 1:2000 dilution of serum (ratio 1:400) in order to nullify the physiological gradient of immunoglobulins across the blood brain barrier (BBB). This procedure allowed a rapid and accurate assessment of the synthesis of specific immunoglobulins in the CSF and a good evaluation of the permeability of the BBB. A diagnosis of SSPE was provided in five out of eight patients with clinical signs of the disease. Clinical follow-up confirmed the diagnosis of SSPE in the group of patients with clear evidence of intrathecal synthesis of antimeasles antibodies.

Isolation of cold-sensitive mutants of measles virus from persistently infected murine neuroblastoma cells.

Clone NS20Y of the mouse neuroblastoma C1300 was infected with wild-type Edmonston measles virus, and, after a transition to a carrier culture, became persistently infected. Persistently infected clones were derived and characterized morphologically by the appearance of multinucleate giant cells and nucleocapsid matrices in cytoplasm and nucleus, but very few budding virus particles. Antimeasles antibodies markedly suppressed the expression of viral antigens and giant cells, and the effect was totally reversible. When the cells were cultured at 33 degrees C, the number of giant cells began to diminish and ultimately disappeared; in contrast, when cultured at 39 degrees C, the cultures invariably lysed. Yields at 33 degrees C were ca. 2 logs lower than those at 39 degrees C. Cells cultured at 33 degrees C produced relatively high levels of interferon, whereas those at 39 degrees C produced little or no interferon. When the persistently infected cultures were exposed to anti-interferon alpha/beta serum at a nonpermissive temperature, there was a marked increase in multinucleate cells, suggesting that maintenance of the persistence state and its regulation by temperature may be related to the production of interferon. Viral isolates from cells cultured at 39 degrees C were obtained, and 90% of viral clones were found to be cold sensitive. Complementation studies with different viral clones indicated that the cold-sensitive defect was probably associated with the same genetic function. Western blot analysis of the persistently infected cells indicated a significant diminution and expression of all measles-specific proteins at a nonpermissive temperature. Infection of NS20Y neuroblastoma cells with the cold-sensitive virus isolates resulted in the development of an immediate persistent infection, whereas infection of Vero or HeLa cells resulted in a characteristic lytic infection, suggesting that the cold-sensitive mutants may be selected or adapted for persistent infection in cells of neural origin.

Immunological Abnormalities and Immunotherapeutic Attempts in Subacute Sclerosing Panencephalitis

This chapter discusses the studies conducted with a large group (13 cases) of patients with subacute sclerosing panencephalitis (SSPE). It presents the results of detailed immunological studies carried out and the outcome of several different attempts at immunotherapy. All 13 of these patients had undisputed SSPE, fulfilling the clinical, virological, electrophysiological and immunological criteria for diagnosis of the disease. One of the patients was first seen before her serum anti-measles antibody titer was increased and when no other features of the disease were present except very mild cerebral involvement that had given rise to persistent nightmares. Six months later a high titer of anti-measles antibody was detected, together with EEG abnormalities. Using the technique of peripheral blood lymphocyte protein synthesis, a subtle but definite lymphocyte abnormality was detected in some of the subjects. There is compelling evidence that humoral immunity is abnormal in SSPE. A possible benefit of thymectomy in this disorder is suggested.

Immunoglobulins and measles antibody in sera of newborn cynomolgus monkeys and in milk of their mothers.

The changes in the serum immunoglobulin (Ig) level and in the measles antibody titer were examined with colony-born infant cynomolgus monkeys from 0 day to 26 weeks in age. In addition, their mother-monkey's milk was analyzed for Ig. The decrease in the IgG level after birth was more remarkable in the infants born with high IgG levels than those born with low IgG levels. The IgM level prominently increased for the first 3 weeks after birth in the infants born with low IgG levels, while the increase was not so prominent in those born with high IgG levels. Anti-measles antibody titers decreased linearly with a half-life of 3.4 +/- 0.4 weeks after birth regardless of the titer at birth. A significant amount of IgA, a very small amount of IgM and little IgG were detected in the milk collected just after parturition. Ig of any class was no longer found in the milk collected one to 6 weeks after parturition. These results suggest that the initial level of transplacentally transferred IgG must be one of the important factors influencing the subsequent development of infant monkey's own immunoglobulin synthesis.

The placental transfer of IgG in the cynomolgus monkey.

The placental transfer of IgG from the mother to her fetus was investigated with colony-bred cynomolgus monkeys. Very low levels of IgG were detected in sera of 84 days old fetus, indicating that transplacental transfer of IgG had started by this fetal age. Afterwards, gradual increase in the IgG level continued till 140 days of fetal age. A marked increase in the level was noted after 140 days of fetal age, during the last 4 weeks of full gestation term. Fetal anti-measles antibody increased in a pattern similar to that in the IgG level. The IgG level and anti-measles and anti-streptolysin O antibody titers of newborns never exceeded those of their mothers.

Antinuclear antibodies in multiple sclerosis.

Low levels of antinuclear antibodies (ANAs) were found by indirect immunofluorescence in the serum of patients with multiple sclerosis (MS). Antibodies were found in 22 (81%) of 27 patients with MS and four (20%) of 20 healthy control subjects, with human epithelial (HEp-2) cells as substrate. Antinuclear antibody titers ranged from 8 to 32 in patients with MS and rarely reached above 8 in control subjects. The most common fluorescence patterns produced by MS serums were diffuse, fine speckled, and diffuse with fine-speckled nuclear fluorescence. Large nucleolar speckles and cytoplasmic fine speckles were also seen. Similar, although stronger patterns were seen in control subjects with positive tests for systemic lupus erythematosus. No correlation was found between ANA and antimeasles antibody titers. The presence of a heterogeneous population of circulating antibodies to a variety of nuclear and cytoplasmic antigens lends further support to the concept that altered immune reactivity exists in MS.

Mechanisms of killing of measles virus-infected cells by human lymphocytes: Interferon associated and unassociated cell-mediated cytotoxicity

Human fibroblasts infected with measles virus express viral hemagglutinin (HA) and fusion (F) glycoproteins on their surface. These infected cells are killed by peripheral blood lymphocytes (PBLs) within 2 to 4 hr of incubation in vitro. Both syngeneic and allogeneic PBLs harvested from human adults, all previously recovered from childhood measles virus infection (immune donors), are efficient in mediating this cytotoxicity. Cytotoxicity is also expressed by PBLs from a donor with no history of measles virus and lacking circulating anti-measles virus antibodies. This cell-mediated cytotoxicity (CMC) is not due to anti-measles virus IgG in that it is not abrogated by purified F(ab′) 2 fragments from rabbit IgG anti-human F(ab′) 2. The killer lymphocytes are Fc-receptor positive and erythrocyte rosetting and non-erythrocyte resetting, as assessed by both positive and negative selection experiments. Exposure of viral glycoproteins at surfaces of infected cells is necessary for this early CMC, which can be completely abrogated by F(ab′) 2 fragments to measles virus glycoproteins. However, the same F(ab′) 2 fragments are not effective in blocking CMC after an 8-hr incubation period (late CMC), suggesting that a different mechanism of CMC operates at the later time. Interferon (IFN)-like activity cannot be demonstrated in culture fluids within 4 hr of incubation, and early CMC is not inhibited by antibody to human IFN. In contrast, after 8 hr of incubation, culture fluids do possess antiviral activity, and do have the capacity to recruit new CMC indicating that IFN is now present in the system. Therefore, viral glycoproteins expressed at the surface of infected cells induced CMC early in the infectious cycle and this CMC is not associated with IFN-like activity. Later during the infection, a different IFN-associated CMC is also present.

Determination of antibody to measles virus by a fluoroimmunoassay (FIAX®)

A fluorometric assay for measuring measles antibody was developed using the FIAX® system. The assay described is simple, practical, and quantitative for the determination of anti-measles antibody in serum. No pre-treatment of any kind is needed prior to assay, and only a single dilution of the patient's serum is needed in order to obtain a titer. Comparative studies of our fluorometric assay and HAI assays show that the two assays provide comparable results in terms of measles antibody titers.

In vitro generation of human cytotoxic lymphocytes by virus. Viral glycoproteins induce nonspecific cell-mediated cytotoxicity without release of interferon.

Purified hemagglutinin and fusion glycoproteins of measles virus either in soluble form or inserted in artifical membranes bind to human peripheral blood lymphocytes and induce cell-mediated cytotoxicity (CMC) in a dose-response fashion. Both autologous and heterologous noninfected target cells are lysed in vitro. The expression of CMC is not inhibited by anti-measles virus antibody added to lymphocytes previously exposed to viral glycoproteins. THe killer lymphocytes are Fc receptor positive, both erythrocyte-rosetting and non-erythrocyte-rosetting, as assessed by both positive and negative selection experiments. The induction of nonspecific CMC by viral glycoproteins either in the soluble state or inserted into artificial membranes could be segregated from the CMC associated with whole virions. First, on kinetics studies, purified viral glycoproteins induced CMC more rapidly than did whole virus. Second, viral glycoprotein-produced response occurred in the absence of detectable release of interferon into the culture medium, whereas CMC activity due to whole virions was associated with interferon release. The fact that purified measles virus glycoproteins integrated into artificial membrane bilayers were as efficient as their soluble counterparts in inducing CMC suggests that the hydrophobic portion of the glycoproteins was not involved in the induction and expression of the lytic activity. Purified glycoproteins from lymphocytic choriomeningitis virus behave similarly, although this virus is unrelated to measles virus. It is inferred that interferon-independent CMC induced by viral glycoproteins might account for some of the biological reactions occurring early in the control of a viral infection.

An acute variant of subacute sclerosing panencephalitis: An autopsy case report

An acute variant of subacute sclerosing panencephalitis (SSPE) was described in a 5-year-old boy who showed rapid progression of coma within 14 days of right hemiplegia. He had measles at 3 years of age. The diagnosis of SSPE was based on the following findings: high anti-measles antibody titer in the serum and in the spinal fluid, periodic complex of EEG, and typical pathological changes of the brain. Treatment with transfer factor failed to improve the worsening clinical course. It is suggested that SSPE should be considered in the differential diagnosis of acute fulminating encephalitides or intracranial vascular lesions.

Lymphocyte-mediated cytotoxicity to measles virus in patients with multiple sclerosis

Peripheral blood lymphocytes from patients with multiple sclerosis were studied for their ability to lyse target cells infected with measles virus using a 51Cr release assay. Immune-specific cytolysis caused by lymphocytes from patients in remission was equivalent to cytolysis observed with lymphocytes from normal immune subjects in both the absence and the presence of added anti-measles virus antibodies. The ability of any individual patient's lymphocytes to kill measles virus-infected targets did not significantly change as the patients' clinical course fluctuated from remission to exacerbation. In addition, sera from multiple sclerosis patients and normal subjects invariably enhanced cytolysis, and no blocking of lymphocyte-mediated killing was observed.

Natural Killing of Measles-Infected Cells by Human Lymphocytes

Abstract HeLa cells persistently infected with measles virus (M-HeLa) show markedly increased susceptibility to natural killing (NK) by normal human blood lymphocytes compared to uninfected HeLa cells. The killing is mediated by the same population of non-B, non-T lymphocytes with avid Fc receptors for IgG that mediate other forms of NK and antibody-dependent cellular cytotoxicity (ADCC). There is no evidence that this NK is mediated by antibody; specifically, the killing regenerates after being abolished by extensive treatment of the effector cells with trypsin or pronase, and it cannot be inhibited by large amounts of F(ab′)2 fragments of anti-measles antibody or by a large excess of normal γ-globulins. In addition, NK of M-HeLa is strongly correlated (r = 0.79, p < 0.001) with NK of an unrelated cell line (K-562) that does not contain measles virus and is a standard target for NK. The strong correlation between NK of M-HeLa and K-562 is confirmed by cold-target inhibition studies showing cross-competition between the two cell lines. Killing cannot be enhanced by addition of supernatants from lymphocytes cultured with M-HeLa to fresh lymphocytes cultured with M-HeLa, HeLa, K-562, or other cell lines, making it unlikely that interferon or other soluble factors have a role in this system. These studies suggest that persistent infection of HeLa cells with measles virus results in the modification of the HeLa cell membrane and that this modification is similar to some structure on K-562 that is important for NK. In addition, these studies raise the possibility that NK could play a role in host defenses against measles virus infection.

Adherence of human peripheral blood lymphocytes to measles-virus infected cells: modulation by solubilized rhesus erythrocyte membranes and carbohydrates.

The adherence of human peripheral blood lymphocytes to HeLa cells persistently infected with measles virus (HeLa-K11) was studied. The following data were observed. (i) The proportion of HeLa-K11 cells with adherent human peripheral blood lymphocytes of rhesus monkey erythrocytes was similar over a wide range of ratios of HeLa-K11 cells to lymphocytes or erythrocytes. (ii) The great majority of human peripheral blood lymphocytes and erythrocytes reacted with the same HeLa-K11 cell (iii). The adherence of lymphocytes or erythrocytes to HeLa-K11 cells was blocked by rabbit anti-measles virus antibody or solubilized monkey erythrocyte membranes. The pretreatment of erythrocytes or lymphocytes with receptor-destroying enzyme did not alter their adherence properties. (iv) The pattern of inhibition observed with several carbohydrates was similar in both the erythrocyte and the lymphocyte adherence assays. These data are consistent with the possibility that the receptor present on both rhesus monkey erythrocytes and human lymphocytes has similar specificities and biochemical composition.

Subacute Sclerosing Panencephalitis

Antimeasles antibody was detected in the saliva of 14 patients with subacute sclerosing panencephalitis. Immunofluorescent staining showed antibody in salvary gland biopsy of 7 patients.

Familial multiple sclerosis: Clinical, histocompatibility, and viral serological studies

Evaluation of presumed "multiple sclerosis families" and comparison with recently reported families has led us to the following observations: (1) Seven of our original fourteen presumptive multiple sclerosis families had to be eliminated after personal clinical evaluation of family members failed to confirm the diagnosis in a second close relative. (2) No segregation of HLA type was noted between affected and unaffected individuals in our seven bona fide multiple sclerosis families, and no consistent segregation was noted in the twenty-eight families reported elsewhere. This supports other genetic evidence that there is not a single, major gene mapping in the HLA complex which predisposes to multiple sclerosis. (3) The DW2 antigen was increased in frequency among affected members of our families, and the A3 B7 haplotype was more frequent among affected members of other families reported. But unaffected members also tended to have an increased frequency of these same antigens. (4) No relationship was noted between HLA type and antimeasles antibody titer within our families.

Immunocytochemical studies for the localisation of measles antigens in multiple sclerosis plaques and measles virus-infected CNS tissue

Actively demyelinating central nervous system (CNS) lesions from a patient with acute multiple sclerosis (MS) were tested for measles antigens using peroxidase-conjugated antimeasles antibody. No evidence of measles antigens was found. Similarly reacted tissue from 2 patients with chronic MS also revealed no evidence of measles antigens. Identically treated and simultaneously tested measles-infected CNS cultures and human SSPE brain tissue stained strongly for measles antigens. The possible reasons underlying the failure to detect measles antigens in MS are discussed.