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OK432 inhibits experimental hepatic metastasis of colon adenocarcinoma ACL-15 in F344 rats.

The effect of OK432 on hepatic metastasis, induced by inoculating 1 x 10(6) ACL-15 cells from a rat colon adenocarcinoma cell line into the ileocolic vein of male F344 rats, was investigated in this study. Metastases were detected 14 days after inoculation in the control rats, however, pretreatment 3 days prior to the tumor cell inoculation with an anti-asialoGM1 antibody, which eliminates natural killer (NK) cell activity in vitro, increased the number of hepatic metastases, shortened the survival time, and decreased the NK activity of the nonparenchymal liver cells (NPC). In contrast, pretreatment with OK432 2 days prior to tumor inoculation significantly decreased the number of hepatic metastases, prolonged the survival time, and augmented the NK activity of the NPC, although treatment with OK432 3 or 7 days after inoculation did not decrease the number of hepatic metastases. Moreover, NPC from the OK432-pretreated rats had a marked antitumor effect against ACL-15 cells in the Winn's neutralization test. The results of this study indicate that pretreatment with OK432 before tumor cell inoculation inhibits hepatic metastasis in this experimental model, possibly by augmentating liver-associated NK activity.

Adenocarcinoma↗

Single and combined infections of specific-pathogen-free chickens with infectious bursal disease virus and an intestinal isolate of reovirus.

The susceptibility of 1-day-old and 7-day-old specific-pathogen-free chickens to infection with a virulent strain of infectious bursal disease virus (IBDV) or an intestinal isolate of avian reovirus, or a combination of the two, was investigated. Chickens infected with IBDV and reovirus had more severe pathological lesions than chickens infected with either virus alone, and prior infection with IBDV enhanced the pathogenicity of enteric reovirus. Virus recovery was attempted from bursa, spleen, thymus, liver, intestine, pancreas, cecal tonsils, heart, and tarso-metatarsal tendons. Viruses were recovered from all tissues sampled for either IBDV or reovirus isolation, and indications were that infection with IBDV before infection with reovirus led to longer persistence of reovirus in some tissues. Antibodies to IBDV or reovirus were measured by the virus neutralization test and enzyme-linked immunosorbent assay. Chickens infected with IBDV had lower (P less than 0.05) antibody responses to reovirus than chickens infected with reovirus alone.

Animals↗

Comparison of measles virus-specific antibodies with antibody-dependent cellular cytotoxicity and neutralizing functions.

Measles virus-specific antibodies with antibody-dependent cellular cytotoxicity (ADCC) and neutralizing functions were compared in 25 healthy women. ADCC antibody was measured in a 12-h 51Cr release assay using peripheral blood mononuclear cells as effectors and Raji cells persistently infected with measles as targets. Neutralizing antibody was determined by the plaque neutralization test (PNT). ADCC and PNT titers correlated well (r = .80, P < .001). ADCC titers ranged from < 10 to > 10(6) and averaged 1.3 logs higher than PNT titers. Three sera with PNT titers of < 120, including 1 with a titer of < 8 (< 10(0.9)), had ADCC titers of > or = 10(2.5). Among subjects born in the United States, ADCC titers, but not PNT titers, correlated inversely with the year of birth, suggesting that ADCC antibody may be more indicative than neutralizing antibody of differences between naturally and vaccine-acquired immunity. These data suggest a possible role for ADCC in protection or recovery from measles.

Adult↗

Virus isolations from mosquitoes collected during the 1982 Japanese encephalitis epidemic in northern Thailand.

From 16 June to 15 August, 1982 CDC light traps were used to collect mosquitoes in the province of Kamphaengphet, N. Thailand. 353,042 mosquitoes comprising 59 species were collected and identified, and 345,173 were placed in pools for attempted virus isolation by inoculation of C6/36 Aedes albopictus mosquito cell cultures. Viruses were isolated from 63 mosquito pools. These comprised 56 flaviviruses, identified as 35 isolates of Japanese encephalitis (JE) virus strains, 18 strains of Tembusu (TEM) virus and three untyped flaviviruses (FLA); three alphaviruses, identified as the first isolates of Getah (GET) virus to have been made in Thailand; and four viruses which are still unidentified. Most virus isolates were from Culex tritaeniorhynchus mosquitoes collected in carbon dioxide baited light traps. JE virus was isolated only over a ten-day period and the last isolate was obtained one week before the peak of admission of human encephalitis cases at Kamphaengphet Provincial Hospital. Rapid screening of isolates grown on Ae. pseudoscutellaris (LSTM-AP-61) mosquito cells by indirect immunofluorescence using flavivirus group-specific and JE-specific monoclonal antibodies showed a high degree of correlation with plaque reduction neutralization tests. An antigen capture enzyme immunoassay (EIA) test successfully identified about 50% of the JE virus positive pools, but the method saved considerable processing time.

Alphavirus↗

Validity of an enzyme-linked immunosorbent assay with serotype-specific monoclonal antibodies for serotyping human rotavirus in stool specimens.

We recently developed a method for serotyping human rotavirus (HRV) by an enzyme-linked immunosorbent assay with HRV serotype-specific neutralizing monoclonal antibodies (ELISA serotyping). In the present study this method was compared with the fluorescent focus neutralization test with serotype-specific rabbit antisera (NT serotyping) in the sensitivity and specificity of the test. Direct serotyping of HRVs which were contained in stool specimens indicated that while only 37% of the samples were successfully serotyped in NT, 78% of the samples could be serotyped in ELISA. Regarding the samples whose serotype could be determined in the two tests, the assigned serotypes were identical in both tests. The results obtained indicated the utility of ELISA serotyping in clinical and epidemiologic studies of HRV infection.

Antibodies, Monoclonal↗

Use of non-neutralizing monoclonal antibodies in an ELISA for intratypic differentiation of 28 echovirus type 25 clinical isolates.

Three non-neutralizing monoclonal antibodies were produced and selected against echovirus type 25 JV-4 prototype strain. They were used in an ELISA to investigate the intratypic differentiation of 28 wild isolates. Clinical isolates fell into seven different groups according to their reactivity patterns in ELISA. Two of the non-neutralizing monoclonal antibodies, 9E4 and 6D3, were highly specific, while the third, 6C9, may recognize an epitope common to other types of echoviruses. In contrast, mouse polyclonal antiserum exhibited large cross-reactivities among echovirus serotypes. The reactivity patterns and the geographical origin of the isolates were generally not correlated and, in the same area, four major antigenic variants sometimes coexisted, especially in the south of France. Moreover, reactivity patterns found with ELISA were hardly ever correlated with those observed in a previous study when neutralization tests were used. These results again underline the non-correlation between structure and biological function in the Picornavirus family.

Animals↗

Observations on the antigenic relationships between Epstein-Barr virus and herpesvirus saimiri.

The antigenic relationships between Epstein-Barr (EB) virus and herpesvirus saimiri (HVS) have been investigated in comparative immunofluorescence, microimmunodiffusion and serum neutralization tests. No similarity was detected between the structural antigens of the two herpesviruses but the results of microimmunodiffusion tests showed that they shared a non-structural, virus-determined antigen. The implications of this finding are discussed in relation to the importance of HVS and its monkey lymphoma as a model system of herpesvirus oncogenesis.

Antigens, Viral↗

Immunity levels against avian encephalomyelitis in vaccinated poultry breeder flocks in Maine.

Four commercial poultry breeder flocks that were vaccinated under field conditions against avian encephalomyelitis (AE) with commercial live or inactivated vaccine were monitored periodically by virus-neutralization testing of blood serum samples and by challenge of their progeny eggs and chicks. The history of the flocks and results of the tests indicate that field exposure might occur during the laying period, thereby boosting immunity titers without causing clinical AE in the progeny chicks. The data also indicate field exposure of certain flocks before AE vaccination.

Animals↗

Antigenic and biological relationships between human coronavirus OC43 and neonatal calf diarrhoea coronavirus.

Monospecific antisera were prepared in mice to human coronavirus OC43 and neonatal calf diarrhoea coronavirus (NCDCV) which had been previously adapted to growth in suckling mouse brain. Brain suspension from infected suckling mice was used as immunogen. The antigenic relationship between OC43 and NCDCV was studied by the indirect immunoperoxidase antibody technique, by the haemagglutination-inhibition (HI) test and a new infectious centre-reduction neutralization test. In mouse immune sera, a two-way cross-reaction between OC43 and NCDCV was detected. However, the antigenic relationship appeared to be closer for internal (as shown by immunoperoxidase staining) as compared to surface antigens (as shown by HI and neutralization). In primary infections of natural hosts there was a high degree of cross-reactivity between the two coronavirus strains for both surface and internal antigens, and homologous and heterologous titres were consistently within an eightfold dilution difference by all tests. Most human adults and calves had antibody to both OC43 and NCDCV and geometric mean titres of homologous antibody were higher than titres of heterologous antibody. Although OC43 and NCDCV share antigenic determinants, they possessed several different biological properties, including plaque morphology by the infectious centre assay, agglutination of 1-day-old chick erythrocytes and resistance of haemagglutinin to physical and chemical treatments.

Antibodies, Viral↗

Development and optimization of plaque assays for rat coronaviruses.

Plaque assays under Sephadex or agarose overlays are described for rat coronaviruses (RCVs) grown in L2 mouse fibroblasts. A plaque assay using Sephadex was simple; however, viable plaques could not be collected for propagation, and fixation was necessary before evaluation. Plaque formation under agarose was optimized using diethylaminoethyl-dextran (DEAE-D) in the pre-treatment and absorption media and trypsin added to the absorption media and agarose overlay. The use of DEAE-D alone, trypsin alone or trypsin combined with DEAE-D significantly increased plaque numbers and visibility. Plaque numbers were highest when pre-treatment media contained DEAE-D, absorption media contained DEAE-D and trypsin, and the agarose overlay contained trypsin. The assay was useful for plaque isolation and quantification of sialodacryoadenitis virus (SDA), Parker's rat coronavirus (PRCV) and other coronavirus isolates from rats and its specificity was demonstrated by plaque-reduction neutralization testing. These methods will facilitate production of cloned virus stocks for study of RCV biology and virus quantification for in vitro and in vivo studies of RCVs.

Animals↗

Teratogenic effects of Colorado tick fever virus in mice.

The teratogenic effects in mice of Colorado tick fever (CTF) virus, a human pathogen, were demonstrated. Various routes of inoculation, gestational periods, and dosages of virus were studied. The incidence of stillbirth and neonatal death increased significantly after inoculation of pregnant mice with CTF virus. CTF virus crossed the placenta and replicated to high titers in the placental and fetal tissues. Mice that survived the first six weeks of life had neutralizing antibody to CTF virus. Neutralization tests, in which pregnant mice were inoculated with both CTF virus and specific antiserum to CTF virus, validated the teratogenic effects of CTF virus in mice.

Animals↗

Serological relationships among feline caliciviruses.

A total of 46 strains of feline calicivirus isolates from the United Kingdom, United States, Australia, and New Zealand were used in an investigation of their serological relationships based on the serum neutralization test. Although demonstrable antigenic variation exists between these isolates, it is shown that significant in vitro cross-activity exists between all these isolates to greater or lesser extent. All isolates tested may be regarded as serological variants of a single serotype of feline calicivirus. It is postulated that this relationship would provide for considerable cross-protection during successive exposures of cats to various feline caliciviruses.

Adsorption↗

Cross-reactive neutralization epitopes on VP3 of human rotavirus: analysis with monoclonal antibodies and antigenic variants.

We analyzed cross-reactive neutralization epitopes on protein VP3 of human rotavirus (HRV) by the use of neutralizing monoclonal antibodies (N-MAbs), which showed a variety of interserotypic reactivity patterns when examined in a neutralization test and an enzyme-linked immunosorbent assay against 15 HRV and 2 animal RV strains. Serological study with the six cross-reactive N-MAbs revealed antigenic variations in some HRV strains within the same serotype as well as a marked antigenic difference between serotype 2 strains and serotype 1, 3, and 4 strains. Epitope analysis of the antigenic variants resistant to the six individual cross-reactive N-MAbs suggested the existence of at least three distinct cross-reactive neutralization epitopes on VP3 of HRV.

Antibodies, Monoclonal↗

Preparation of Hybridomas producing monoclonal antibodies against human interferon.

To prepare hybridomas secreting monoclonal antibodies (MoAb) against human alpha-interferon (alpha-IFN), BALB/c mice were immunized with IFN produced in Namalwa cells. Native alpha-IFN, as well as partially purified or on cellulose adsorbed alpha-IFN preparations were used for immunization. Seven hybridomas continuously secreting IgG against human alpha-IFN were prepared by fusion of splenocytes from immunized donors with the mouse myeloma cells. MoAb reacted in ELISA as well as in neutralization test with human lymphoblastoid, leukocytic and recombinant alpha-IFN.

Animals↗

Multiple recombinant ELISA for the detection of bovine viral diarrhoea virus antibodies in cattle sera.

The most immunogenic proteins (E0, E2 and NS3) of bovine viral diarrhoea virus (BVDV) (NADL strain) were expressed in the baculovirus/insect cells system. Recombinant antigens were applied to the design of enzyme immunoabsorbent assays (ELISAs) for the detection of specific antibodies in cattle sera. The assays developed were shown to be highly sensitive and specific in comparison with the viral neutralization test, which is the reference test for the serological diagnosis of BVDV. The present results demonstrate the contribution of each recombinant antigen to determine clearly the pattern of anti-BVDV antibodies in bovine serum samples.

Animals↗

Serologic classification of two ovine adenovirus isolates from the central United States.

Two ovine adenovirus (OAV) strains (RTS-42 and RTS-151), isolated from lambs in the central United States, were compared using 2-way cross-neutralization tests with the 6 recognized OAV species, 9 bovine adenovirus species, and 4 porcine species. Virus RTS-42 was identified as OAV type 5, confirming previous results. Virus RTS-151 was identified as OAV type 6, although the serologic crossing was largely one-sided.

Adenoviridae↗

Fluorescent antibody responses of cases and contacts of hand, foot, and mouth disease.

Paired sera from clinical cases, familial contacts, and school contacts of hand, foot, and mouth disease were tested by indirect immunofluorescence against a strain of coxsackie A16 virus which had been adapted to human fibroblast tissue-cultured cells. All of the 22 clinical cases developed immunoglobulin (Ig) G antibodies, and 11 responded with IgM and 10 with IgA antibodies. Seventeen of 21 familial contacts of cases showed subclinical infection as determined by development of IgG antibodies. Only seven and nine, respectively, of these had demonstrable IgM and IgA antibodies. Thirteen of 16 school classmates of three clinical cases were shown to have experienced subclinical infection by the development of IgG antibodies. Only five had antibodies in the IgM fraction, and three had antibodies in the IgA fraction. A comparison of IgG titers with those obtained by neutralization tests provides further evidence that the indirect fluorescent antibody technique represents a rapid diagnostic procedure for this disease.

Adolescent↗