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[Passive immunization against rubella: studies on the effectiveness of rubella-immunoglobulin after intranasal infection with rubella vaccination virus].

The effectiveness of high-titre rubella immunoglobulin was tested on rubella-susceptible female juveniles or young adults after intranasal immunization with rubella strain RA 27/3, 20 ml of rubella immunoglobulin having been administered intramuscularly to 56 subjects at different times (1, 3 and 5 days) after the immunization. The effect was demonstrated by seroconversion and virus isolation from the throat. In the control subjects (26) the secroconversion was 96% and in 42% of subjects virus was demonstrated in the throat. Early administration of rubella immunoglobulin (up to three days after immunization) depressed the seroconversion rate to 55% and virus isolation rate to 17.5%. Later administration (five days after immunization) increased the conversion rate to 81%, the isolation rate to 31%. There were no serious side effects of the immunization and the injection of the immunoglobulin. The results indicate that on early administration of an adequate amount of high-titre rubella immunoglobulin a protective effect can be expected, although this favourable effect-obtained in the conditions of this study-cannot be unreservedly transposed to the situation in wild virus infections.

Adolescent

Pathogenesis of the rubella exanthem: distribution of rubella virus in the skin during rubella with and without rash.

In a previous assessment of the role of rubella virus in the pathogenesis of the rubella exanthem, virus was consistently isolated from cell cultures of skin biopsy specimens of the rash, and it was concluded that presence of virus in the skin was essential to evolution of the rash. For determination of whether virus is present in the skin only in association with rash, punch biopsies were performed concurrently on areas of skin with and without rash. Among paired skin specimens of 16 patients, virus was isolated from sites of rash in 12 and from the uninvolved skin in 10. In another patient, shown by serologic response and recovery of virus from the pharynx to have rubella without a rash, virus was also isolated from the skin. It is concluded that rubella virus is widely disseminated in the skin of patients with rubella irrespective of the presence or distribution of the rash, and that the presence of virus in the skin, although a constant feature of the disease, is only one of the factors involved in the pathogenesis of the exanthem.

Biopsy

Persistent rubella-specific IgM reactivity in the absence of recent primary rubella and rubella reinfection.

Between two and seven sera from cases of persistent detection of rubella-specific IgM for periods in excess of 2.5 months, but in the absence of recent primary rubella or rubella reinfection, were examined for rheumatoid factor, heterophile antibody, and IgM reactivity against toxoplasma and a number of viruses. The relative avidity of the rubella-specific IgG1 has been assessed in all the sera by two methods. None of the sera contained rheumatoid factor or heterophile antibody, nor did any contain detectable concentrations of IgM specific for any of the panel of antigens apart from five sera which contained low concentrations of IgM specific for some coxsackieviruses B. No sera were positive for low avidity specific IgG1 although three did give equivocal results with one avidity test and one gave equivocal results with the second avidity test.

Animals

Cell-mediated immunity in rubella assayed by cytotoxicity of supernatants from rubella virus-stimulated human lymphocyte cultures.

Rubella virus-stimulated lymphocytes from rubella-seropositive donors produced in the culture medium cytotoxic activity with preferential action against rubella-infected over uninfected target cells. The ability of lymphocytes to produce the cytotoxic activity upon stimulation by rubella virus correlated with the humoral rubella-immunity status, i.e. no such cytotoxic activity developed in the supernatants of lymphocyte cultures of rubella-seronegative donors. Stimulation of lymphocytes from seropositive donors by rubella virus was also detected by thymidine incorporation, but the correlation of lymphocyte responsiveness to the humoral rubella antibody status was not so clear as in the cytotoxicity assay. Conversion of lymphocytes from unresponsive to responsive to rubella virus following natural rubella infection and after rubella vaccination was demonstrated using both methods. Following vaccination rubella-specific cell-mediated immunity first became demonstrable at 14 days. The responsiveness of lymphocytes to phytohaemagglutinin (PHA) after rubella vaccination was followed by studying thymidine uptake and the ability of lymphocytes to produce lymphootoxin. By both tests marked suppression of PHA response occurred at days 3 and 7 after vaccination.

Adult

[The role of rubella specific IgM within the serological diagnosis of rubella (German measles) (authors transl)].

The sera of 327 patients were investigated for rubella specific IgA after separation of the IgM fraction by sucrose density gradient centrifugation. 1. In 96 out of 137 cases suspicious of clinical rubella, where the time for demonstration of a significant rise of the antibody titer had been missed, the demonstration of rubella specific IgM confirmed the diagnosis of German measles. This demonstration was still possible in 9 out of 10 cases 5 to 7 weeks after the onset of the disease, but in only 2 out 7 cases 8 to 15 weeks after the onset of the disease. 2. In 48 patients who had contact with patients with German measles but did not develop any symptoms, a primary infection could not be excluded by the examination of the whole serum. In 5 out of them rubella specific IgM indicted a recent rubella infection. 3. Rubella specific IgM could not be demonstrated in 47 patients who had high antibody titers but no other hints for a recent rubella infection. 4. In 89 newborns, the investigation was prompted, when elevated IgM levels were found in umbilical cord blood, or when the newborn was sick, or when a rubella infection of the mother was suspected or proven. Rubella infection of the newborn was confirmed by rubella specific IgM only if the mother had contracted German measles during the first trimester of the pregnancy (6 out of 7 cases). The diagnosis of primary rubella infection during pregnancy is proven only if 2 out the fose in antibody titer, demonstration of rubella specific IgM.

Antibodies, Viral

Cytotoxic activity against rubella-infected cells in the supernatants of human lymphocyte cultures stimulated by rubella virus.

Supernatant fluids of lymphocyte cultures from rubella-seropositive donors, stimulated with inactivated rubella virus, showed cytotoxic activity against rubella-infected target cells (NYU 32 line of human embryonic fibroblasts) but not against uninfected fibroblasts. The time of appearance of cytotoxic activity in rubella-stimulated lymphocyte cultures correlated with increased rate of DNA synthesis as measured by thymidine uptake. No such cytotoxic activity became detectable in the supernatants of lymphocyte cultures from rubella-seronegative donors cultured in the presence of rubella virus, or in unstimulated lymphocyte cultures from seropositive or seronegative donors. The cytotoxic activity was lost at 60degreesC in 30 min. In contrast to this rubella virus-induced cytotoxic activity, cytotoxin produced in mitogen-stimulated lymphocyte cultures from rubella seropositive and seronegative donors was equally cytocidal against rubella-infected and uninfected human fibroblasts. Although the nature of cytotoxic activity remains to be characterized, it is suggested that it is associated with a lymphokine released immune-specifically from rubella virus-stimulated lymphocytes.

Antibodies, Viral

[Diagnosis of rubella by demonstrating rubella-specific 19 S and 7 S antibodies (author's transl)].

Identification and measurement of rubella antibody of 19 S (IgM) and 7 S (IgG) immunoglobulins by sucrose density gradient centrifugation and a modified hemagglutination-inhibition (HAI)-test is described. Human group 0 of red blood cells were employed after trypsination. The method proved to be highly reliable and sensitive for the rapid detection of rubella IgM antibody in routine research to make an accurate diagnosis of recent rubella infection. Sera from 185 pregnant women, 200 children suspect of congenital infection, 15 mothers of intrauterin infected children, and 32 cases of rubella vaccine-induced immunity control were studied. In postnatal rubella infections, specific IgM-antibody could be detected in all sera after the onset of the rash up to eight or ten weeks after infection. From March 1973 until September 1974 laboratory diagnosis confirmed 30 of 185 pregnant women to have been recently infected with rubella. In less than 3% (4 cases) the results could not confirm or exclude a recent history of rubella. Among 200 children with congenital defects or some irregular development, specific IgM antibody was detected in 36 cases. The diagnosis of congenital rubella was confirmed or detected in the first months of life. One of the infected children was well at birth but later developed mental retardation and hearing loss, and some others showed only minor defects. In some children rubell IgM antibody persisted 5-14 months. IgM rubella antibody was not found in the sera of 15 mothers from children with intrauterine infection confirmed by IgM antibody demonstration. Post-vaccination-tests showed serum IgM and IgG immunoglobulin-responses to be similar to the natural rubella-virus-infection. Maximal titers of both immunoglobulin-fractions were but four-fold lower after vaccination (28 of 32 tested cases). The described method is able to verify the time of infection by testing a single serum. The HAI-test of the 12 fractions collected from each serum after density-gradient centrifugation shows a typical titer-profil for every time of immunization status: beginning with traces of IgM-antibody - when the HAI-test of the uncentrifugated serum may be less than 1:10 - until 10 weeks after infection when only IgG-antibody will be detectable. The greatest amount of IgM-antibody is found in the 2. to 4. week after the rash.

Adult

Rubella, measles and mumps antibodies following vaccination of children. A potential rubella problem.

One hundred sixty-eight children immunized by one suburban Minneapolis clinic during routine pediatric visits had serum antibodies measured to determine the efficacy of rubella (HPV77 DE5 strain), measles (Edmonston B and Moraten strains), and mumps (Jeryl Lynn strain) vaccines. Serologic failure rates at the mean postvaccination times tested were as follows: rubella, 36% (4.7 years); measles, 18% (6.5 years); and mumps, 9% (4.5 years). Antibody titers shortly after vaccination were not done, so seronegative subjects may never have responded or their titers may have declined with time; our rubella data suggest the former. Children vaccinated with rubella and measles at less than 14 months of age had higher failure rates than those vaccinated at a later age. This supports postponement of rubella and measles vaccinations until at least 15 months of age. In addition to current measles reimmunization policies, consideration also should be given to reimmunizing girls who were given rubella vaccine at less than 14 months of age. Twenty-four percent (19/79) of children vaccinated with HPV77 DE5 strain rubella at 14 months or older had rubella hemagglutination-inhibiting titers less than 8. This is disturbing and, if confirmed by others, would prompt the use of a different strain of rubella vaccine for routine immunization.

Adolescent

[Rubella in pregnancy: experimental studies on the value of gamma-globulin after rubella wild virus infection (author's transl)].

Rubella infection was produced with a wild rubella virus in 24 volunteers by intranasal application. Some of the subjects were given gamma-globulin intramuscular 24 hours later. In the control group clinically manifest rubella with massive viral excretion in the throat occurred in all in whom infection had taken place. In the majority of subjects rubella virus could be demonstrated in blood shortly before the skin rash appeared. High antibody titres occurred already 20 days after the infection. In subjects who were given gamma-globulin there were no clinical signs of rubella, viral excretion in the throat was diminished and shortened and viraemia was never demonstrated, antibody response was delayed and lower. But there was no significant decrease in the infection rate after gamma-globluin administration. It is concluded that early administration of a sufficient amount of rubella antibodies achieves inhibition of rubella virus multiplication and thus decreases significantly the risk of damage to the fetus.

Adult

Lymphocyte responses to rubella antigen and phytohemagglutinin after administration of the RA 27/3 strain of live attenuated rubella vaccine.

Lymphocyte phytohemagglutinin (PHA) responsiveness was found suppressed in both rubella sero-negative and sero-positive recipients of RA 27/3 strain of live attenuated rubella vaccine; the suppression was readily demonstrable only when a suboptimal dose of PHA was applied in the test. Lymphocytes from sero-negative vaccinees, which initially showed little or no in vitro response to concentrated rubella virus, became responsive after vaccination by day 21, when the highest sensitization to rubella antigen was seen. In the sero-positive vaccinees. lymphocytes responded to rubella antigen in vitro before vaccination, and in most cases vaccination did not result in significant changes in lymphocyte response. These results suggest that rubella vaccination leads to temporarily increased lymphocyte reactivity to rubella antigen, and the increased lymphocyte response to specific antigen may occur at the time of mild suppression of PHA response.

Adult

Immunosuppression and isolation of rubella virus from human lymphocytes after vaccination with two rubella vaccines.

Two groups of young rubella-susceptible women were vaccinated with two rubella vaccines. Heparinized blood samples were taken from all individuals the day of vaccination and 5, 7, 15, 21, 30, 35, and 42 days later. Purified lymphocytes from these samples were cocultivated with AGMK cells for rubella virus isolation. Parallel samples of lymphocytes were stimulated with phytohemagglutinin, and the rate of [14C]thymidine incorporation was determined. Rubella virus was isolated from lymphocytes collected on days 7, 15, and 21 after RA27/3 vaccination in contrast to days 7 to 35 after HPV77 vaccination. The lymphocyte response to phytohemagglutinin was markedly suppressed from day 5 to 15. Normal lymphocyte responses were restored within 1 month after vaccination with RA27/3, but even later (1 week) after HPV77 vaccine. Lymphocytes from rubella-susceptible persons infected invitro with rubella virus vaccines and stimulated with phytohemagglutin displayed a decrease in their responsiveness to the mitogen similar to that observed with lymphocytes from vaccinees. The transient immunosuppression observed in vaccinees is probably due to virus-induced functional damage of the lymphocytes since no direct cytocidal effect of rubella vaccine has been demonstrated on human lymphocytes.

Adult

Diagnosis of postnatal rubella by the enzyme-linked immunosorbent assay for rubella IgM and IgG antibodies.

A semi-automated enzyme-linked immunosorbent assay (ELISA) was established for the detection of rubella IgM antibodies in non-fractionated sera. A cut-off level between rubella IgM positive and negative sera was determined by a study of sera without rheumatoid factor from 200 blood donors. Testing of 12 donor sera containing rheumatoid factor showed that 5 sera gave a positive result in the rubella IgM assay. Rubella IgM antibodies were quantified by ELISA in a study of 214 serial serum specimens drawn from 16 patients with rubella during a period of up to 10 years after the infection. Peak values of the IgM antibodies were reached approximately 8 days after onset of the rash, and the persistence of the IgM antibodies ranged from 17-90 days, with the exception of one patient with a prolonged IgM response. The rubella IgG antibodies increased slowly after the rash and reached maximum levels about 50-120 days, after which a monor decrease was observed. The results of the present study indicate that ELISA is suitable as a routine procedure for the serodiagnosis of recent rubella.

Acute Disease

Rubella in Jerusalem. 1. Seroepidemiologic findings in children born after the 1972 rubella epidemic.

Nine hundred and sixty-nine babies born after a severe rubella epidemic were tested at the age of eight months for the presence of hemagglutination inhibition antibodies to the disease. There were 208 (21.9%) seropositive babies with titers of greater than or equal to 1:16. Seven cases of congenital rubella were diagnosed at birth, 14 were confirmed by eight months and two babies, who were considered normal at the age of eight months, developed late signs of the disease. Less than 3% of the mothers of the seropositive babies had had clinically recognized rubella in pregnancy. If positive hemagglutination inhibition antibody to rubella accurately reflects congenitally acquired rubella infection, between 8.1 and 21.9% of the 969 babies had serologic evidence of the disease. For cases of congenital rubella diagnosed by the age of three years, the limits are between 8.9 and 23.7 per 1,000 live births for the risk of the disease at the height of the epidemic. Rubella vaccine was not available in Israel until after the 1972 epidemic, the effects of which justify the continued search for and immunization of seronegative women of childbearing age, in addition to routine immunization of young girls.

Adult

Absence of cell-mediated immunity to rubella virus 5 years after rubella vaccination.

The long-term effectiveness of rubella vaccination in childhood is particularly important because the ultimate goal of immunization is the prevention of infection during pregnancy. Of 25 healthy children tested 4 to 5 years after rubella vaccination, 19 showed no evidence of cell-mediated immunity (CM) to rubella virus despite the presence of hemagglutination-inhibition or complement-fixation antibodies or both. Twenty-two of 25 seropositive, naturally infected young adults showed evidence of CMI. These results indicate that fetuses of women who have been vaccinated against rubella may not be protected against damage by wild rubella infection during the pregnancy, when CMI is physiologically depressed.

Adult

Parity of women contracting rubella in pregnancy. Implications with respect to rubella vaccination.

Data from the National Congenital Rubella Surveillance Programme showed that 44% of children with congenital rubella reported to the programme were born to primiparae. This high proportion is thought to be due to the fact that there was a two-fold increase in the rate of abortion for rubella in pregnancy for women with two or more children. This higher incidence of congenital rubella in firstborns emphasises the need for rubella vaccination prior to a woman's first pregnancy.

Abortion, Therapeutic

Single-radial-hemolysis test for the assay of rubella antibody in antenatal, vaccinated, and rubella virus-infected patients.

The single-radial-hemolysis test (SRHT) is a simple, convenient, and sensitive method that provides accurate measurement of IgG antibody to rubella virus. Lipoprotein inhibitors of rubella hemagglutination do not interfere with the SRHT; thus the assay of low levels of antibody is more reliable by SRHT than by the rubella hemagglutination-inhibition test (RHIT). The technique for the SRHT in a form suitable for large numbers of tests is detailed and compared with that of the RHIT, and the reproducibility of the SRHT in routine operation is assessed. Results obtained by testing various populations in Western Australia are used to show the application of the SRHT in the diagnosis of rubella and in the monitoring of rubella vaccination programs.

Adolescent

Rubella in Jerusalem. 2. Clinical and serologic findings in children with congenital rubella.

Forty-eight children born with clinical and serologic manifestations of congenital rubella were followed for a three-year period. Expanded rubella syndrome, multiple anomalies and single defects were found, mostly in the child's first year of life. Some new organic problems were found at a later age in children who had initially been healthy but who had been followed up because of their high antibody levels to rubella. Of the 48 children, 15 (31%) were born to mothers who had had clinically and serologically diagnosed rubella during pregnancy. In 33 children (69%) the mother had had asymptomatic rubella. These findings emphasize the need to identify and immunize seronegative women before pregnancy.

Abnormalities, Multiple