HI Antibody Research Articles

抗ＨＩ抗体による寒冷凝集素症にＩＴＰを合併したＥｖａｎｓ症候群の１例

A 70-year-old female was admitted with anemia and thrombocytopenia. Her blood type was B Rho (+). Her RBC was 250×104/μl, Hb7.8g/dl, Ht 23.2%, Retic. 55‰, WBC 2900/μl, T. bil 0.3mg/dl, LDH 917U/L, and Haptoglobin <10mg/dl. Cold agglutinin titer was×1024, direct Coombs test (-), indirect Coombs test (4+). Antibody screening test in saline revealed serum agglutinated adult O cells (4+) (×512), adult i cells (+) (×64), cord cells (-) and B cells (-), leading to an antibody diagnosis of anti-HI antibody. She subsequently developed thrombocytopenia (16×103/μl) and PAIgG was 54.8ng/107 platelets, giving a diagnosis of idiopathic thrombocytopenic purpura together with cold agglutin disease. The presence of cold agglutinin disease due to anti-HI antibody and development of ITP indicated this to be a case of Evans syndrome. Hemolytic anemia due to anti HI antibody is very rare, as is the association of ITP with cold agglutination disease.

Dose-dependent antibody response to influenza H1N1 vaccine component in elderly nursing home patients.

The effects of an increased antigen dose on HI, IgG, IgA, and IgM antibody responses to influenza A/Taiwan/1/86 (H1N1) were investigated in 92 elderly nursing-home residents (mean age 81 years) and 104 young subjects (mean age 20 years). At a standard 10-microg dose, HI and IgG titer rises were lower in the elderly. HI titers did not improve at higher vaccine dosages. By contrast, influenza-specific IgG and IgA antibody responses were dose dependent in elderly subjects, but not in young. In the young subjects, IgM antibody responses were dose dependent. The improved antibody responses in the elderly as observed in IgG and IgA were not reflected in the HI response. Therefore, the evaluation of antibody production by HI only may lead to an underestimate of the immune response in elderly people.

Immunogenicity of subunit trivalent influenza vaccine in children with acute lymphoblastic leukemia.

The aim of this study was to assess humoral response to influenza vaccine in children with acute lymphoblastic leukemia. Studies were performed in 25 patients previously vaccinated against influenza (Group A) and in 20 children who had never been immunized before (Group B). In Autumn, 1996, they were vaccinated with subunit trivalent influenza vaccine containing 15 microg of hemagglutinin of A/Singapore/6/86, A/Wuhan/359/95 and B/Beijing(184/93. Antihemagglutinin (HI) and antineuraminidase antibody titers were determined before immunization and 3 weeks and 6 months after vaccination by the hemagglutinin inhibition test and the neuraminidase inhibition test. All results were presented as the geometric mean titer of antibodies, mean fold increase of antibody titer, protection rate and response rate. In Group A mean fold increase of HI antibodies ranged from 17.2 to 26.7 three weeks after vaccination and from 22.1 to 38.2 six months after vaccination, while in Group B it ranged from 15.7 to 22.6 and from 30.3 to 39.3, respectively. In the case of neuraminidase, mean fold increases for Group A varied from 9.2 to 13.2 three weeks after immunization and from 15.6 to 21.1 six months after vaccination, whereas for Group B they varied from 5.5 to 8.3 and from 14.4 to 23.4, respectively. Six months after vaccination the proportion of subjects with HI antibodies > or = 1:40, as well as those with at least 4-fold increase of HI antibody titers, ranged from 68 to 100% in Group A and from 90 to 100% in Group B. No vaccinated child was infected with the influenza virus; the vaccine was well-tolerated and did not cause any adverse reactions. The results obtained in this study indicate that influenza vaccine is immunogenic in patients with acute lymphoblastic leukemia, despite their serious disease.

Antibody response of inactivated influenza vaccine in the elderly

The antibody responses of 31 elderly persons (11 males and 20 females, mean 82.2 years) and 23 adults (mean 40.7 years) to inactivated influenza vaccine were studied. Participants were given twice the influenza vaccine containing A/Yamagata/32/89, A/Kitakyusyu/159/93, and B/ Mie/1/93. Serum specimens were obtained before vaccination, and 2 weeks and 3 months after the second vaccination. Antibody to influenza was determined by hemagglutination inhibition test (HI). The rates of fourfold or greater increases of HI antibody against the A/Yamagata, A/ Kitakyusyu, and B/Mie were 96.8%, 100%, and 74.2%, and there was no significant difference of the rates of fourfold or greater increases between elderly persons and adults. The rates of possession of HI antibody more than 128 in titer after vaccination were significantly higher than those of HI antibody before vaccination, and were maintained at high levels up to three months after the second vaccination. These results suggest that antibody response to the inactivated influenza vaccine is satisfactory in the elderly.

Hemagglutination with crane herpesvirus.

A crane herpesvirus (CrHV) grown in chicken embryo fibroblast (CEF) cells was tested for hemagglutination (HA) with erythrocytes from a variety of species at 4 degrees C, room temperature and 37 degrees C. HA was observed at all temperatures with erythrocytes from mouse, ddY and BALB/c strains, but not with those from cattle, sheep and chicken. Mice, ddY strain, showed an individual variation in agglutinability of their erythrocytes and erythrocytes from BALB/c gave a higher HA titer. The HA activity was inhibited by the sera obtained from naturally infected cranes, experimentally infected duck and immunized rabbit with CrHV. HI antibody titers of these sera showed a closely positive correlation with their neutralizing antibody titers.

Seroepidemiological survey for influenza virus in Belém, Pará, Brazil, 1992-1993

A seroepidemiological study was carried out from 1992 to 1993 to estimate the prevalence of antibody to prevailing influenza virus strains circulating among patients attending the IEC Virology Laboratory in Belem, Northern Brazil. A total of 179 (11%) of serum samples were obtained during the post-epidemic period and processed by the hemagglutination inhibition test against the A/Taiwan/1/86 (H1N1), A/Beijing/353/89 (H3N2), and B/Yamagata/16/88 types of influenza virus. The serological results indicate circulation of viruses antigenically related to all three strains during the two-year study period. In 1992, the overall prevalence rates of HI antibodies to H1N1 and H3N2 were 84% and 56%, respectively. Presence of antibodies to both strains was detected in all age groups, suggesting intense viral activity that year. Antibodies to influenza B were detected at lower levels in all patients during this period. Similar prevalence to the H1N1 virus was noted in 1993, indicating that this strain occurred in both years. However, an increase in prevalence rates for the H3N2 virus was seen in 1993, suggesting that this strain (or a related virus) circulated intensively during this year. Influenza B activity also increased in 1993, causing infection mainly among young adults.

Estimation of antibodies to Newcastle disease virus in tears.

Micro-haemagglutination inhibition tests (Micro-HI) were used to measure the level of maternal IgG in the tears of chicks and also to measure the levels of HI antibodies in the tears and serum after vaccination with "F" strain of Newcastle disease virus (NDV) and in the face of an outbreak of Newcastle disease. There was a 1.4 fold difference between the maternal IgG concentration in the serum and tears. The ratio of serum IgG to lachrymal IgG after maternal transfer was 4 to 5:1 on day 4 to 9 and decreased to 2.6:1 on day 12 post-hatch. The intra-ocular vaccination of chicks with "F" strain of NDV resulted in the highest titre of HI antibodies in the tears though there was no significant difference in the response of chicks vaccinated through intranasal, oral and intravenous routes. In the face of an ND outbreak, the level of HI antibodies in the tears during the acute phase was very high and persisted at the same level for 14 days.

Detection of PMV‐1 specific antibodies with a monoclonal antibody blocking enzyme‐linked immunosorbent assay

A highly reproducible monoclonal antibody (Mab) blocking ELISA (B-ELISA) has been developed and evaluated for the detection of NDV-specific antibodies. The Mab utilised is specific for a conserved PMV-1 serotype-specific epitope, as demonstrated by the indirect immunoperoxidase test. It reacted with all strains representing different serogroups within the PMV-1 serotype, but not with any strain belonging to other PMV serotypes. Sensitivity and specificity of the B-ELISA were compared with the haemagglutina-tion inhibition test (HI). Blocking and HI antibodies were detected in sera of chickens 8 days post-experimental infection. The B-ELISA proved consistently more sensitive than the HI test. In another survey, 62 sera from experimentally vaccinated chickens were tested; 95.2% proved positive by B-ELISA, 85.5% by indirect ELISA and 74% by HI test. When 504 field sera from vaccinated chickens and turkeys were tested, 98% were positive by B-ELISA, and 69% by HI. The specificity was evaluated by testing 1066 samples from NDV-free flocks, all of which proved negative by both methods. Other advantages of the B-ELISA include easy standardization and quality control, and ability to test sera from any species (including exotic or wild birds as well as mammals). The use of low dilution serum or egg-yolk samples makes the test quick and easy to perform and suitable for large-scale screening.

Diagnosis of respiratory virus infections using GACELISA of urinary antibodies

Sensitive and specific methods are needed to diagnose respiratory virus infections using body fluids such as urine that, unlike blood samples, are readily obtained by non-invasive means. Immunoglobulin G antibody capture enzyme-linked immunosorbent assays were developed for detection of antibody rises to respiratory syncytial virus and influenza A/Taiwan 3H1N1) after initial quantification and adjustment of urinary IgG concentration. Of 24 elderly subjects whose sera were assayed by the complement fixation test for antibody to RSV, seven had convalescent titres ≥ 32, and five had ≥ 4-fold rises in titre. Acute and convalescent urines for six of these seven subjects were tested for virus-specific urinary IgG by GACELISA. Four of four persons with ≥ 4-fold rises in CFT had urine ELISA convalescent to acute ratios of ≥ 1.8 whereas two subjects with convalescent CF titres > 16, but no increase in serum antibody titre, had urine convalescent/acute ratios of 1.0. Ten subjects with ≥ 4-fold rises in CFT or HI antibodies to influenza A/Taiwan had urine ELISA ratios of ≥ 1.4 when samples taken on the day of influenza vaccination and 16 days later were compared. These preliminary observations demonstrate clinically significant rises in respiratory pathogen antibody levels between acute and convalescent urine samples, provided that total urinary IgG concentrations are quantified and then standardised.

Laboratory virological diagnosis of imported dengue cases

Suspected dengue cases in Japan during the period of 1985-1995, 173 cases with unknown febrile illness entering or returning from mostly southeast Asia, were tested by serology and RT-PCR gene amplification. Seventy seven cases were confirmed by a significant rise of dengue 2 (Den 2) HI antibody in paired sera or by detection of HI antibody titer higher than 1:320 in single serum. The other 3 cases with antibody levels not higher than 1:80 in paired sera and 12 cases with an antibody 1:160 in single sera were positively suspected of dengue infection but were not confirmed. Countries of origin of confirmed cases were Thailand (39 cases), Philippinse (15), India (13), and Indonesia (9). Sera of dengue cases showed high degrees of cross reactivity of Japanese encephalitis virus (JEV) in HI test but not in IgM capture ELISA. Sera of confirmed JEV-infected cases, however, showed practically no cross reactivity to Den 1 4 in HI test, suggesting unilateral cross reactivity of HI antibody. RT-PCR detected the Den 1 genome in sera of 3 cases obtained within 3 days after onset and the Den 2 genome in serum of case 4 days after onset. Although the number is limited, 92 (53%) out of 173 cases of febrile illness of unknown etiology entering Japan from tropical countries were either confirmed or positively suspected to be dengue fever. Considering possibilities of under reporting, importations of tropical viral infections should be bigger in number and will necessitate our intensified alertness.

Hemagglutination with porcine reproductive and respiratory syndrome virus.

Porcine reproductive and respiratory syndrome (PRRS) virus grown on MARC-145 cell cultures was tested for hemagglutination (HA) with erythrocytes from a variety of species at 4 degrees C, room temperature and 37 degrees C. HA was observed at all temperatures with mouse erythrocytes but not with cattle, sheep, goat, horse, swine, guinea pig, mongolian gerbil, goose and chicken erythrocytes. The HA activity was enhanced by treatment of virus materials with Tween 80 followed by treatment with ether. The HA titer and HA pattern of virus materials treated with Tween 80 and ether (TE) were 4- to 8-fold higher and more clear than those of the virus materials without TE treatment. The optimum conditions consisted in pretreatment of virus material with Tween 80 at a final concentration of 0.06-0.125% (v/ v) for 15-60 min followed by treatment with ether at a concentration of 50% (v/v) for 5-15 min in ice bath with continuous shaking. The curve of active virus production in intra- and extracellular virus samples resembled that of HA production although it rose somewhat earlier in intracellular virus samples. The HA reaction was inhibited by specific antiserum. HI antibody titers of individual pig sera showed a significant positive correlation with their neutralizing antibody titers.

Prevention of influenza by the intranasal administration of cold-recombinant, live-attenuated influenza virus vaccine: importance of interferon-γ production and local IgA response

To clarify which immunological factors were more effective in preventing influenza virus infection, we measured immunological parameters induced by vaccination and infection in vivo and in vitro. Healthy adult subjects (n = 128) were divided into vaccinated (n = 85) and untreated (n = 43) groups. Eighty-five were vaccinated intranasally with a trivalent cold-adapted recombinant influenza virus vaccine containing type A (H1N1 and H3N2) and B viruses. Subjects were mostly seropositive before vaccination. In 29 (80.6%) of the 36 examinees showing a prevaccination HI antibody titre of less than 1:128, the titre increased more than four times after vaccination. On the other hand, an increase of more than four times was found in four (8.2%) of the 49 individuals who had shown a prevaccination titre of more than 1:128. The IgA antibody was negligibly detected in the nasal wash specimens before vaccination, and was induced by vaccination in some cases. Lymphocyte proliferation and interleukin 2 (IL-2) production in cultured lymphocytes of the same subjects stimulated by H1N1 virus in vitro were correlated with the HI antibody titre. However, the interferon γ (IFN-γ) production was low before vaccination, regardless of the HI antibody titre, and showed a significant increase after vaccination. It was suggested that local IgA response and IFN-γ production play important roles in the prevention of influenza. Since there was the outbreak of influenza A (H1N1) in Kochi Prefecture after completion of blood samples 6–8 weeks after the second vaccination, we examined the above hypothesis. A significantly (p<0.01) lower morbidity rate in the vaccinated examinees was found. The HI antibody, lymphocyte DNA synthesis and IL-2 production, which had been increased by vaccination, showed little further increase in response to the epidemic. In contrast, local IgA response and IFN-γ production, which had been increased by vaccination, showed another increase because of the epidemc.

冬期における豚血中からの日本脳炎ウイルスゲノムの検出

By reverse transcription-coupled polymerase chain reaction with nested primerse, the RNA genome of Japanese encephalitis virus (JEV) was detected from 23 of 371 swine blood specimens collected in Kagawa prefecture from June 1993 to May 1994. The genome was detected from 8 of 161 specimens collected during a winter period from January to April in 1994. HI antibody was detectable throughout the year and 2 specimens collected in January were positive for IgM antibody, but no infectious virus was isolated during the winter time.

Inducement of respiratory mucosal immunogenicity for nasal vaccine due to amine- and amide-inactivation of parainfluenza virus type 1, Sendai.

The protective effects in mice by nasal vaccination of amine- and amide-inactivated Sendai viruses were investigated by a contact exposure experiment, immunofluorescent examination of the entire respiratory tract, and checking the serum HI antibody development. Of 10 monoamines, ethanolamine and 2-methoxyethylamine vaccines induced complete protection, and methylamine, ethylamine, n-propylamine, n-butylamine, 2-ethoxyethylamine, diethylamine and triethylamine vaccines brought about almost complete protection or lower respiratory infection. The methoxyamine-treated mouse conferred the least protection. Of 5 diamines, 1,3-diaminopropane vaccine inhibited completely the infection, but hydrazine, ethylenediamine, putrescine, and cadaverine vaccines produced regional infection. Two polyamines, spermine and spermidine, did not inactivate the virus. Of 4 amides, only semicarbazide vaccine conferred complete mucosal defense, while acetamide, propionamide, and isonicotinic acid hydrazide vaccines lead to regional infection. Serum HI titers developed by vaccination were low on the whole, following their slight rise, fall or maintenance postexposure. In effect, the 4 vaccines inactivated by a best-suited interstrand cross-link between phosphate groups in helix of viral RNA brought about the strongest protection, and showed the necessity of a definite length of molecules for inactivants.

Interaction between chicken anaemia virus and live Newcastle disease vaccine

Three groups of 150 SPF chickens were spray-vaccinated with live Newcastle disease La Sota-type vaccine (clone 30) at one day of age, and another three groups were NDV spray-vaccinated at 10 days of age. In each of the two series of NDV-vaccinated groups, one group also received at day-old 10(5) TCID50 of chicken anaemia virus (CAV) also and another group 10(5) TCID50 of CAV plus a low dose of virulent Marek's disease virus (MDV). After one week, chickens of the groups which had been NDV-vaccinated and CAV-infected at day-old, with or without MDV, showed severe respiratory distress, conjunctivitis, drooping wings and ruffled feathers. After two weeks, wet and inflamed eyes were observed. After three weeks the respiratory problems were overcome, but the entire group showed retarded growth as compared with the group which had received NDV vaccine only. The 'respiratory sounds' were milder in the chickens NDV-vaccinated at 10 days of age, about 10% of the chickens showing retarded growth. Mortality in CAV-infected chickens which had received NDV vaccine at day-old was above 30% at 4 weeks of age, and between 15 and 20% when NDV had been administered at the age of 10 days, and was 5% in the two NDC vaccine control groups. Decreased haematocrit levels were measured in all four CAV-infected groups at 14 days of age. In serum samples collected for 6 weeks at weekly intervals from chickens of the six groups, no differences were observed between HI antibody titres against NDV virus. Thus, dual infection with CAV and live NDV vaccine did not impair the humoral immune response against attenuated Newcastle disease vaccine.

Radioimmunoprecipitation and immunoblot studies of antibodies to rubella virus in patients with chronic liver disease.

Patients with autoimmune chronic active hepatitis (AICAH) and some other chronic liver disorders often have very high titres of rubella HI antibodies. In the present study sera from 46 patients with chronic liver disease and controls were examined for rubella antibodies using radioimmunoprecipitation assay (RIPA) and Western blot. RIPA appeared to be more suitable than Western blot for the study of the individual antibody specificities provided that proteins (possibly actin) interfering with the resolution of the E2 glycoprotein band are identified. It was shown that patients with high rubella HI titres reacted strongly against the E1 glycoprotein and in general also against the core protein (C). Reactivity to the E2 glycoprotein was detected with all sera from patients with chronic liver disease but varied more in strength. Three patients with post-acute rubella showed very faint E2 reactivity, but strong E1 and C reactivities. Patients with primary biliary cirrhosis had normal HI titres and showed no increase in reactivity in RIPA. The present findings show that patients with chronic liver disease and high rubella HI antibody titres exhibit an enhanced specific antibody response to rubella virus structural proteins.

EDS 76 HI antibodies in a flock of free-range layers

An investigation of poor laying performance in a flock of free-range hens revealed high levels of serum antibodies to EDS 76 in the flock initially examined and in another, older flock on the same farm. These flocks had contact with ducks on a farm dam and were supplied with untreated drinking water from the dam. Serological evidence indicated that another flock supplying the same egg packing station had been infected with EDS 76 virus. Little serological evidence of EDS infection was detected from five other flocks supplying the packing station, parent breeders or the ducks resident on the dam. Therefore, the source of the EDS 76 virus remains conjectural.

Effects of Polychlorinated Biphenyls on Humoral Immunity and Blood Serum Vitamin A and E Concentration of Chicks

Halouzka R., V. Jurajda and E. Minksova: Effects of Polychlorinated Biphenyls on Humorallmmu­ nity and Blood Serum VitaminA and E Concentration o/Chich. Acta vet. Bmo, 63, 1994: 141-144. The effects of ingestion of a feed contaminated with polychlorinated biphenyls (PCB) using Delor 103, a technical preparation, on the dynamics ofhaemagglutination-inhibiting (lfl) antibo­ dies to Newcastle disease virus (NOV) after administration of Czechoslovak: vaccine A vipest (LaSo­ ta) were studied in an in-vivo experiment. Blood serum vitamin A and E concentrations were asses­ sed at the end of the experiment. Brown Hisex layer-type hibrid cockerels from a commercial flock were fed a feed mixture supp­ lemented with Delor 103 at the rate of 25 mg.kg-I or 50 mg.kg-I feed from 1 day of age till the end of the experiment in the 10th week. In the 3rd post-hatching week they received vaccine against NDV in drinking water. The level of post-vaccination immunity was checked at 14-day intervals. Vitamin concentrations were assessed with fluorescence spectrophotometry. Compared with the control birds, the two PCB-fed groups showed a lower HI antibody level in the 4th, 6th and 10th week. No differences in the reduction of immunity due to the different PCB levels in the feed were recorded. Vitamin A concentration was significantly (P:s 0.05) lower in both experimental groups than in the controls. Vitamin E concentration of the group fed the feed mixture contaminated with Delor 103 at 25 mg.kg-I was significantly (P :s 0.05) higher than in the controls. Continued feeding of a PCB-contaminated feed mixture to chickens may be involved in delayed development, fluctuation or reduction of their post-vaccination humoral immunity against NDV. Brown Hisex chickens. PCB, post-vaccination immunity. NDV. Atrophy of lymphoid tissues observed in various animaJ species after ingestion of feeds containing polychlori­ nated biphenyls (PCB) has been regarded as a morphological correlate of their immunosuppressive effects (C 0 s 1972; Vos andDe Roij 1972; Vos andvan Driel-Grootenhuis 1972;Halouzka andJurajda 1991). However, published direct evidence for depressed immune response in poultry has been scarce. In a previous report (H al 0 u z k a et al. 1990) we demonstrated morphological changes in the lymphoid organs of chickens fed a PCB-contaminated feed. The present report was designed to study the effects of PCB on the deve­ lopment and dynamics of humoral antibodies after vaccination against Newcastle disease and on blood serum vita­ min A and E concentration.

Recycling of H1N1 influenza A virus in man--a haemagglutinin antibody study.

Sera from people born between 1883 and 1930 and collected in 1977 were tested for the presence of HI antibodies to A/FM/1/47 (H1N1) virus and three recently (1977 and 1978) isolated influenza A-H1N1 viruses. The highest frequency of high-titred antibody to the four H1N1 viruses was detected in sera from people born in 1903-4, i.e. 42, 54, 38, and 22% had antibody against A/FM/1/47, A/Hong Kong/117/77, A/Brazil/11/78, and A/Fukushima/103/78 respectively. The birthdate groups 1896-1907 showed a higher percentage of HI antibody titres greater than or equal to 18, greater than or equal to 50, greater than or equal to 100 or greater than or equal to 1600 against the four H1N1 viruses than the birthdate groups 1907-30. This indicates the existence of an era, 1908-18, in which, apart from the H3N2 virus (1900-18), the H1N1 virus was epidemic among the human population.

日本の軽種馬におけるウマインフルエンザウイルスの3株のワクチン株に対するHI抗体保有状況

日本の軽種馬におけるウマインフルエンザウイルスの3株のワクチン株に対するHI抗体保有状況