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In hepatitis B virus (HBV) infection-preventing programs, serum or plasma levels of antibody to hepatitis B surface antigen (anti-HBs) are important to determine whether individuals are protective or not. We compared anti-HBs responses using passive hemagglutination assay (Mycell) and chemiluminescent immunoassay (Architect) among four recipient groups of HB vaccines, Meinyu, HBY, Bimmugen and Heptavax II, that have been approved in Japan. Overall, in a total of 1875 vaccinees Mycell results showed recipient groups of Meinyu and HBY acquired higher anti-HBs levels than those of Bimmugen and Heptavax II. Comparison of anti-HBs responses by both Mycell and Architect in recipient groups of Meinyu (n=150), HBY (n=218), Bimmugen (n=260), and Heptavax II (n=47) demonstrated the order of vaccinees' responses, such as geometric mean titers, ratios of acquiring high antibody levels (Mycell titers over 1024, Architect measurements over 1000 mIU/mL), and ratios of having unsuccessful antibody responses (Mycell titers under 8, Architect measurements under 10 mIU/mL), were somewhat different between the two assays. Comparison of Architect measurements at given Mycell titers revealed Bimmugen-recipients showed significantly lower values than HBY- or Heptavax II-recipients. Around critical protective levels, 5 of 22 Bimmugen-recipients with Mycell titers 16 or 32 showed Architect measurements under 10 mIU/mL, while 8 of 11 Heptavax II-recipients with Mycell titers below 8 demonstrated Architect measurements over 10 mIU/mL. Thus, discrepancies in anti-HBs evaluation between Mycell and Architect seemed to partly depend on administered vaccines. These results indicate anti-HBs concentration should be evaluated carefully so that we could completely prevent HBV infection.

In order to know the rubella immune status of Japanese women aged 20 to 39 years old, we analyzed the hemagglutination-inhibiting antibody titration data on 264,371 sera which were sent to a commercial diagnostic laboratory from gynecology clinics all over Japan during 1999 through 2003. We found that antibody-positive rates remained at about 95% during the period, but, from 2000, geometric mean antibody titers of the positive sera gradually decreased each year. Annual analysis of the data will be useful for monitoring the trend of rubella immunity among Japanese women of childbearing age.

Our previous study found mistakes by some doctors in the choice of an assay for determining antibodies in Japan. To compare the positivity rates for antibodies by assays, we measured the antibodies of measles, rubella, chickenpox and mumps from the same sera using such methods as the EIA, HI and CF assays. The subjects were 175 nursing students. The positivity rates for measles, chickenpox and mumps by the EIA assay were 96.6%, 93.7%, and 83.3%, respectively. Those for rubella by the HI and CF assays were 92.0% and 10.1%. The sensitivity rates for measles, chickenpox and mumps by the HI and CF assays, based on the results of the EIA assay, were 75.1%, 102.4% and 69.2% in the HI assay, and 20.6%, 38.7% and 8.0% in the CF assay, respectively. Our previous study showed that the sensitivity of the HI assay for rubella antibody is same as that of the EIA assay in Japan. Currently an EIA assay should be chosen for these antibodies and the HI assay or IAHA assay should be possible selections for rubella and chickenpox. However, international comparison of the cutoff titers for these antibodies should be considered.

Among 888 patients who underwent operation or endoscopic resection for gastric cancer (1994-1998), 75 patients, who had no colorectal disease or only small polyps 5 mm or less in diameter, were positive on the immunologic fecal occult blood test (IFOBT) (the positive group). They are compared with the other 813 patients (the negative group) as to the following 6 points: symptoms, presence of anemia, depth of invasion including macroscopic appearance, location, maximum diameter of lesions, and microscopic findings. The rate of positive-IFOBT gastric cancer was 8.4%. The average blood hemoglobin concentration was significantly lower in the positive group than in the negative group. Advanced cancers, especially type 2 and 3, were significantly more frequent in the positive group than in the negative group. The size of the lesions tended to be larger in the positive group than in the negative group. There was no difference between the groups as to symptoms, location, depth of invasion and microscopic findings. In conclusion, IFOBT-positive patients who have no colorectal disease or only small polyps 5 mm or less in diameter should be recommended to undergo upper gastrointestinal endoscopy.

We analyzed the anti-HIV antibody titer by particle agglutination (PA) on 11 HIV seroconversion panels. PA titer increased very rapidly and the titer went up to 1,000 or more within 18.2 days after seroconversion. The results suggest that one to two weeks of duration will be enough to differentiate persons at the early stage of HIV infection from individuals with HIV screening test initially reactive but false positive. In Japan, HIV prevalence is very low and the majority of the HIV screening test-positive (reactive) cases turned out to be false positive. This HIV testing strategy (one to two weeks interval bleeding) will be very practical and useful to differentiate early stage of HIV infection cases from the majority of false positive cases.

To investigate sensitivity to rubella virus (RV) in healthy individuals, we examined levels of antibodies to RV in sera by an indirect immunofluorescence assay (IFA) and compared levels of antibodies by IFA with those by a hemagglutination inhibition (HI) assay. Of 114 healthy individuals, we detected antibodies to RV in serum specimens from 103 (90.3%) by IFA and in those from 109 (95.6%) by HI assay. The peak value of levels of antibodies by HI assay was 4 fold higher than that by IFA. When levels of antibodies by IFA were less than 32, levels of antibodies by HI assay ranged from < 8 to 1024. We did not detect anti-rubella antibodies of IgM class in all serum specimens and detected anti-rubella antibodies of IgA class in serum from only 1 individual by IFA. We detected antibodies to rubella in sera from 51 (94.4%) by IFA and in sera from 52 (96.3%) by HI assay of 54 individuals who reported having had rubella, and in sera from 23 (88.5%), by IFA and in sera from 26 (100%) by HI assay of 26 individuals reported having been vaccinated. Also, we detected anti-rubella antibodies in sera from 13 (76.5%) by IFA and in sera from 15 (88.2%) by HI of 17 individuals who reported having had neither rubella nor vaccination. In serum from 1 individual who reported having had rubella, we detected antibodies to rubella by IFA but not by HI assay. In serum specimens from 2 individuals who reported having had rubella vaccination, from 3 having had vaccination, from 2 having had neither rubella nor vaccination, we detected anti-rubella antibodies by HI assay but not by IFA. On the other hand, by both assays, we detected antibodies to RV in all sera of individuals who reported having had rubella and been vaccinated. The serodiagnosis, at least, by two methods is necessary to prevent individuals from rubella virus infection, because of following results: 1) influence of an inhibitor in serum specimens was thought to be variable. 2) The results measured by IFA were differed from those by HI assay in some individuals. 3) It is difficult in diagnosis of rubella from clinical symptoms alone. Also, it might be required to use vaccine to the individual who lacks detectable antibodies to rubella in serum by any method to prevent rubella infection.

An antigenicity study of (+/-)-4-diethylamino-1,1-dimethylbut-2-yn-1-yl 2-cyclohexyl-2-hydroxy-2-phenylacetate monohydrochloride monohydrate (NS-21), a new drug for the treatment of urinary frequency and incontinence, was conducted in Hartley guinea pigs and BALB/cAnN mice. The following results were obtained. No active systemic anaphylaxis reactions were found in guinea pigs immunized by subcutaneous injection of NS-21 alone or in combination with Freund's complete adjuvant (FCA). No 24-hr heterologous passive cutaneous anaphylaxis reactions were elicited in rats by sera from mice immunized by intraperitoneal injection of NS-21 alone or in combination with 3% aluminum hydroxide gel. No passive hemagglutination reactions were elicited by sera from mice immunized by subcutaneous injection of NS-21 in combination with FCA. These results show that NS-21 has no antigenicity under the present experimental conditions.

The antigenicity was tested of a new antineoplastic agent S-1 (a combination of tegafur (FT), CDHP and potassium oxonate (Oxo)) in mice and guinea pigs. 1. Male BALB/c or C3H/He mice were sensitized with S-1, CDHP, Oxo, and conjugates of CDHP (or Oxo) and human serum albumin (HSA). S-1 was administered by oral gavage, and the other compounds were administered intraperitoneally with adjuvant (alum). In the heterologous passive cutaneous anaphylaxis (PCA) test, using Sprague-Dawley rats as recipients, no IgE antibodies against S-1, CDHP, or Oxo were detected to any serum obtained from the sensitized mice, and no eliciting antigenicities were seen for CDHP or Oxo. 2. Male Hartley guinea pigs were sensitized with S-1, CDHP, Oxo, and conjugates of CDHP (or Oxo) and HSA. S-1 was administered by oral gavage, and the other compounds were administered subcutaneously with Freund's complete adjuvant (FCA). The homologous PCA test, active systemic anaphylaxis test, and passive hemagglutination test showed no production of antibodies against S-1, CDHP, or Oxo in any sensitized guinea pig, and no eliciting antigenicities for CDHP or Oxo. 3. Female Hartley guinea pigs were sensitized with S-1 subcutaneously with FCA. The active cutaneous anaphylaxis test revealed that S-1 did not induce cell-mediated delayed type hypersensitivity. 4. These results indicated that S-1, Oxo, and CDHP were not antigenic in mice and guinea pigs.

The practicability of four diagnostic methods for entamoebiasis histolytica including micro-gel diffusion precipitin test (MGDP), indirect immunofluorescent antibody test (IFA), indirect hemagglutination test (IHA) and enzyme-linked immunosorbent assay (ELISA) was evaluated. The serological test methods were compared by using sera obtained from 30 entamoebiasis histolytica patients and 130 normal health individuals. The highest sensitivity was obtained with the method of ELISA, followed by IFA and IHA : the lowest was obtained with MGDP. On the contrary, the high specificity was obtained with IHA, IFA, MGDP : the lowest was obtained with ELISA. Intensity of the antibody titers in IHA was correlated well with that of IFA. In addition, we studied antibodies nonspecifically reactive to Entamoeba histolytica in sera from E. histolytica-negative individuals with high CRP patients with regard to the sensitivity and specificity. Among 101 sera examined, six showed false positive results of which five were the sera with extremely high CRP.

Antigenicity studies of T-3761, a new quinolone derivative, were conducted and the following results were obtained. 1. By active systemic anaphylaxis test in guinea pigs, neither immunogenicity nor allergenicity of T-3761 was noted. 2. By homologous 4-hour passive cutaneous anaphylaxis (PCA) test using the serum obtained from these guinea pigs, neither immunogenicity nor allergenicity of T-3761 was noted. 3. The potential of IgE antibody production in mice was examined by heterologous 24-hour PCA test in rats. But neither the potential of IgE antibody production nor allergenicity of T-3761 was noted. 4. By passive hemagglutination assay, we analysed the antibody titer in rats and dogs serum obtained from three-month toxicity repeated dose studies (oral and intravenously administration). But the hemagglutination response was negative and specific antibodies were not detected.

A new immunochromatography assay (Dainascreen Ausab Dainabot) has been recently introduced for the detection of the presence of antibody to HBsAg. To evaluate the feasibility of using the Dainascreen Ausab, we carried out comparison tests with this method and PHA. In the test of 439 sera from HB vaccinees, inhabitants in Iki Island, Nagasaki Pref., patients with autoimmune diseases and with acute hepatitis B, 154 (31.2%) were positive by Dainascreen Ausab, 145 (29.4%) were positive by PHA and 145 (29.4%) were positive by both Dainascreen Ausab and PHA. Nine (1.8%) were positive by only Dainascreen and there were none positive by only PHA. A good correlation was observed between the titer of the antibody by this method and IMx. The anti-HBs assay by this method was able to be completed within 15 minutes and the procedure was very simple. The results indicate that the sensitivity of Dainascreen is superior to PHA and that it is easy to use.

In order to evaluate the feasibility for the detection of antibody to hepatitis C (anti-HCV), the first generation assay (c100-3 Ab) and two second generation assays (2nd EIA and 2nd PHA) were used to test 477 individuals who visited the medical hospital or clinics in Iki Island, Nagasaki Prefecture. HCV RNA, antibody titer by 2nd PHA and four kinds of antibody to epitope of HCV by RIBA II were also surveyed to determine their association with these three assays. Prevalence of anti-HCV was 26.6% by c100-3, 38.8% by 2nd PHA and 39.6% by 2nd EIA, indications that the 2nd generation assays are much more sensitive than c100-3. Prevalence of HCV RNA was 82.1% among 190, anti-HCV positive individuals; 100% among 52 individuals with liver disease, but only 75.4% in those without liver disease. HCV antibody titer, over 2(11) was higher among those who were positive for HCV RNA than those negative for HCV RNA. Four antibodies by RIBA II were all positive and reacted strongly when they were positive for HCV RNA, but only antibody to core antigen was observed among those negative for HCV RNA, suggesting that only antibody to core antigen remains in those with past HCV infection.

Lactitol hydrate (lactitol) was tested for its antigenicity in guinea pigs and mice. The following results were obtained. 1. No active systemic anaphylaxis reactions were found in guinea pigs sensitized subcutaneously with lactitol alone or in combination with Freund's complete adjuvant (FCA). 2. No 24 hr heterologous passive cutaneous anaphylaxis reactions were elicited in rats by sera from mice sensitized intraperitoneally with lactitol alone or in combination with 3% aluminum hydroxide gel. 3. No passive hemagglutination reactions were elicited by sera from mice sensitized subcutaneously with lactitol in combination with FCA. From these results, it is concluded that lactitol has no antigenicity under the present experimental conditions.

A questionnaire survey of blood donors at Santa Cruz General Hospital, Bolivia, showed that while there were no seropositive cases of syphilis, HBsAg, and HIV, the prevalence of Chagas' disease was very high (23%) among the 225 blood donors who responded to this questionnaire. Actual cases of Chagasic seropositive blood being used for blood transfusion were seen, including the urgent need for a program for Chagas' disease in Bolivia. From the results of this study, it is recommended that for blood donors from South American countries, the presence/absence of Chagas' disease should be confirmed.

A new immunochromatography assay technique (DAINASCREEN HBsAg, Dainabot) has been recently introduced for the detection of the presence of hepatitis B surface antigen (HBsAg). To evaluate the feasibility of using the DAINASCREEN HBsAg, we carried out comparison tests with this method and RPHA. Results obtained were as follows: In a test of 692 sera from inhabitants in Iki island, Nagasaki Prefecture, 24 (3.5%) were positive for HBsAg by both DAINA and RPHA. Twenty-four patients with hepatitis B were all positive in both methods. Thirty patients with autoimmune diseases and eight who had accidental needle exposure to the blood of patients with HBV infection were negative by both methods. Six individuals were followed over a period of 10 years and all eventually eliminated HBsAg from their sera. In four patients, HBsAg remained positive by DAINA at one yearly check after it had become negative by RPHA. In five patients, HBsAg remained positive by RIA and IMs after DAINA and RPHA had become negative. A good correlation was observed between DAINA and RPHA with a concordance rate of 99.5%. A good correlation of HBsAg titer between the two methods was observed and found to be significant (r = 0.87, p < 0.001). HBsAg assay was able to be completed within 15 minutes by DAINA and the procedure was simple. These results indicate that the sensitivity of DAINA is superior to RPHA and it is easy to use.

Sixty four cases of autoimmune hemolytic anemia (AIHA) referred to our laboratory from 1985 to 1993 consisted of 51 warm type AIHA, 9 cold agglutinin disease (CAD), one paroxysmal cold hemoglobinuria, and 3 mixed type AIHA. There were 5 patients who had all clinical features of AIHA except for a positive direct-antiglobulin test (DAT). These patients were diagnosed as DAT-negative AIHA because of the elevation of red blood cell-associated IgG (RBC-IgG). The mean RBC-IgG of 100 healthy individuals was 33 +/- 13 (SD) molecules per one RBC. On the other hand, the RBC-IgG of patients with DAT-positive and with DAT-negative warm type AIHA were ranged from 257 to 12,421 and from 126 to 256 molecules per one RBC, respectively. One of 9 patients with CAD had a hemolytic anemia associated with only a cold agglutinin titer of 32 using saline-suspended RBCs, but a titer of 4,096 in the presence of bovine albumin. This case was diagnosed as low titer CAD. All of patients with DAT-negative AIHA or low titer CAD showed a good response to corticosteroid treatment.

An EIA for measuring anti-TPO autoantibodies (rhTPO-EIA) was developed using recombinant human TPO expressed in CHO cells and was compared with MC-HA generally used in laboratory routine work. rhTPO-EIA showed a satisfactory reproducibility in the intra-assay test and did not have an accidental error of lots. Almost equal number of healthy females and males were measured for their IgG binding to TPO to define a normal range of anti-TPO autoantibodies. After setting 20 IU/ml as an upper limit of normal range, sera from patient with thyroid disorders were measured for their anti-TPO autoantibodies. Chronic thyroiditis and Graves' disease were highly positive, while adenoma, thyroid cancer, SLE, and RA were low in their positivity. The positive rate of anti-TPO autoantibodies was compatible to those of previous reports in each disorder. Seventy-two sera from patients with chronic thyroiditis or Graves' disease were measured for their autoantibodies by both rhTPO-EIA and MC-HA and the results were compared between both methods. A correlation coefficient was 0.486. Following absorption with thyroglobulin, sera were measured again and as the results, the correlation coefficient increased to 0.723. Therefore, MC-HA was thought to be influenced in the presence of anti-thyroglobulin autoantibodies. Since rhTPO-EIA is excellent in quality and not affected by anti-thyroglobulin antibodies, it is useful and applicable to clinical diagnosis and observation of thyroid disorders.

Herpes simplex virus type 1 (HSV-1) has been recently known to agglutinate mouse erythrocytes. We performed the hemagglutination (HA) test under various conditions to investigate the nature of hemagglutinin of HSV-1. HA of mouse erythrocytes was observed with glycoprotein C (gC)-positive HSV-1 strains but not with gC-negative HSV-1 mutants and HSV-2 strains. A recombinant HSV-1 constructed by transfecting a gC-negative strain with a gC-1 gene regained full HA activity. The erythrocytes from several strains of rats as well as mice were agglutinated by gC-positive HSV-1 strains. Formalin fixation of erythrocytes and ultraviolet inactivation of virus did not reduce the HA activity. The HA activity which was inhibited by heparin and dextran sulfate appeared late in the process of virus multiplication and was inhibited by adding cytosine arabinoside or tunicamycin to the medium. The HA activity was inhibited not only by antisera against HSV-1 but by antisera against HSV-2, although their hemagglutination inhibition antibody titers were low. These date support the importance of gC in HSV-1 and heparan sulfate moieties at the surface of erythrocytes as previously reported. HA by HSV-1 might provide a new serological diagnostic test for HSV infection.

Treponema pallidum subsp. pallidum Nichols (Tp) antigens were purified by centrifugation in the presence of Hypaque (sodium diatrizoate) or MgCl2. When the TPHA (Treponema pallidum hemagglutination) tests with crude and purified antigens were carried out in the sera of patients with untreated primary syphilis, the purification by MgCl2 resulted in enhanced sensitivity to Tp-specific IgM as the same as the purification by Hypaque. In addition, the SPHA (solid phase hemadsorption) test with the Hypaque-purified antigen to anti-Tp-IgM antibodies was carried out using the sera of patients with primary and secondary syphilis. Fifty-nine percent (27/46) of them showed positive for the crude antigen and 100% (46/46) for the purified antigen, indicating that sensitivity to anti Tp-IgM was clearly enhanced with the antigen purification. The captured Tp-IgM-RIA test with the purified antigen was also undertaken in 16 patients with primary syphilis, 31 patients with secondary syphilis and 73 patients with latent syphilis. All the patients in the former two groups were positive, while all of the patients in the latter group were negative. To elucidate the mechanism whereby the purification improves the sensitivity to anti-Tp-IgM antibodies, host-derived antibodies associated with crude and purified Tp cells were measured by a fluorescent antibody technique. As the results, a great amount of rabbit anti-Tp-IgM antibodies were found on the surface of the crude antigen, whereas only a trace amount was detected on the purified antigens.(ABSTRACT TRUNCATED AT 250 WORDS)

The incidence of P-fimbriated E. coli from patients with pyelonephritis, cystitis and asymptomatic bacteriuria was 78.6%, 31.9% and 22.2%, respectively. Almost all of the P-fimbriated E. coli have also type-1 fimbriae. In the in vitro test, P-fimbriated E. coli attached to the uroepithelial cells in higher number than the type 1 fimbriated E. coli. The results of the adhesion inhibition test suggested that simultaneous presence of P-and type 1 fimbriae is the most significant virulence factor in urinary tract infections.

The present study was carried out using a low-molecular antigen to clarify which of the two IgG subclasses (IgG1 and IgG2) of guinea pigs is responsible for the immunological assay systems in ordinary antigenicity tests for the development of a novel drug, and to what extent such IgG subclass antibodies contribute to the assay systems in addition to IgM antibody. Guinea pigs were immunized with TNBS plus FCA (3 mg/body), at intervals of 10 days, 3 times in total. The anti-TNBS serum was fractionated into peak I (IgG2), peak II (IgG1) and peak IV (IgM) by DEAE-cellulose column chromatography, and the immunological activities as well as the functions of the above three peaks were estimated by HA, homologous PCA reaction, PSANA and P-Arthus. The IgM and two IgG peaks possessed immunological activities on HA and P-Arthus, and the following degree of activities was shown in both assay systems: peak II (IgG1) greater than peak IV (IgM) greater than peak I (IgG2). In the homologous PCA reaction and PSANA, immunological activities were seen only in peak II (IgG1). It is confirmed that the IgG1 subclass is a homocytotropic antibody involved in the PCA and PSANA assay systems in the case of low-molecular immunogens such as TNBS.

In recent years, a great advance has been brought in an early detection of colorectal cancer, especially early colorectal cancer. But some controversies are present as follows: 1. Mass screening for colorectal cancer. Immunologic tests for fecal occult blood increased the sensitivity and specificity for detecting colorectal neoplasia without a question. However, this screening method has not proven to get the effectiveness in decreasing mortality of colorectal cancer, although a case control study is under way in our department. 2. Adenoma-carcinoma sequence or de nov carcinoma. These two theories on histogenesis of colorectal cancer are on "hot" discussion to be better understood. 3. Strip biopsy for colorectal lesions. This method is now begun to be introduced to apply for small type-II lesions of colon. 4. Others (barium enema or endoscopy, a new method of colonic preparation which replaces Brown's method, tumor marker in colorectal cancer etc.). Today's status and future prospects were made on these matters.

In the management of a chorionic disease, human chorionic gonadotropin (hCG) is the most reliable tumor marker. However, hCG has a low titer, so that it cannot be strictly distinguished from the human luteinizing hormone (hLH) by the traditional method that uses a hemagglutinin reaction (HAR) or by a radioimmunoassay (RIA). Recently, however, the detection of the beta-COOH-terminal peptide of hCG (hCG-CTP) by an enzyme immunoassay has made it possible to clearly distinguish hCG from hLH, and from April, 1987 to December, 1989, 13 trophoblastic diseases have been managed using this new technique. Results have shown that human chorionic gonadotropin-CTP, when compared to other methods of measurement, is the most sensitive tumor marker and as it is the most accurate, it should be used in careful follow-up observations of a chorionic disease.

Immunological properties of cefpirome sulfate (CPR) were examined. The immunogenicity and challenging ability of CPR were examined in guinea pigs by active systemic anaphylaxis (ASA) and homologous 4-hr passive cutaneous anaphylaxis (PCA) tests. The animals given CPR alone intraperitoneally for sensitization and their sera were negative for ASA or PCA reactions, like the results with reference substances, ceftazidime (CAZ) and cephalothin sodium (CET). When each antibiotic plus Freund's complete adjuvant (FCA) was used for sensitization, ASA reactions were observed with CPR, cephaloridine (CER), CET, and cefazolin sodium (CEZ), and PCA reactions, with CPR and CET. CPR had the ability to challenge the ASA and PCA reactions. CER and CET also showed the ability to challenge ASA or PCA reactions, though at low incidences. The cross-reactivity of CPR with commercially available antibiotics was examined by heterologous PCA test and by passive hemagglutination test and its inhibition test. The antiserum used was from rabbits immunized with each antibiotic-ovalbumin conjugate plus FCA, and the antigen was each antibiotic-bovine serum albumin conjugate. CPR cross-reacted markedly with cefotaxime sodium (CTX) having the same side chain at position 7 and showed weak, unidirectional reactions with CAZ and CET. In the in vitro direct Coombs test, the positive reactions noted with CPR were stronger than those with latamoxef sodium, equal to those with CEZ and slighter than those with CTX, CET and benzylpenicillin potassium. In conclusion, in the safety evaluation of CPR, its antigenic potential may not be a problem, like the cases of other antibiotics.

Immunogenicity, eliciting antigenicity and cross-reactivity of new cephem antibiotics, BMY-28100 and cefepime, were studied by means of passive cutaneous anaphylaxis (PCA), passive hemagglutination (PHA) and active systemic anaphylaxis in guinea pigs, and of PCA in mice and the results were compared with those obtained with reference antibiotics. In addition, the direct Coombs' reaction of the human blood was examined in vitro for the test antibiotics as compared with reference antibiotics. The results obtained are summarized as follows: 1. Immunogenicity Immunogenicity of unconjugated antibiotics was examined using the corresponding conjugates with bovine gamma-globulin (BGG) as eliciting antigen. When used as emulsions with Freund's complete adjuvant, cephalothin (CET) and benzylpenicillin (PCG) produced IgG1 and IgM antibodies in guinea pigs. However, cefepime as well as cephalexin (CEX) did not produce these antibodies, and BMY-28100 showed slightly active sensitization only for anaphylactic shock. In BALB/c and C3H/He mice, BMY-28100 and cefepime failed to produce antibodies under the experimental condition while IgE antibody formation to CET was observed. 2. Eliciting antigenicity Unconjugated CET and PCG provoked anaphylactic signs in guinea pigs sensitized with their conjugates with rabbit serum albumin (RSA). Cefepime, however, provoked no anaphylactic shock and BMY-28100 as well as CEX showed slight signs. In the other systems examined, no reactions were observed when elicited with BMY-28100, cefepime or the reference antibiotics. 3. Immunological cross-reactivity BMY-28100 did not cross-react with the reference antibiotics. While the antiserum to the RSA conjugate of CET provoked weak cross-reaction on PHA with the BGG conjugate of cefepime, the antiserum to the RSA conjugate of cefepime failed to react with the BGG conjugate of CET. Other cross-reactivities of cefepime were not observed against the reference antibiotics. 4. In vitro direct Coombs' reaction BMY-28100 did not induce the Coombs' reaction of the human blood in vitro at the testable concentration of 10 mg/ml. Cefepime or cefazolin (CEZ) caused no reaction even at a high concentration of 80 mg/ml, while CET and PCG caused a positive reaction at 10-40 mg/ml and 60 mg/ml, respectively. As shown above, immunogenicity and eliciting antigenicity of BMY-28100 and cefepime were somewhat weaker than CET and PCG but similar to CEX, and cross-reactivities of the test antibiotics with these reference antibiotics were not observed in general. The ability of BMY-28100 to give a positive reaction in the Coombs' test was weaker than that of CET, and that of cefepime was weaker than CET and PCG and equivalent to CEZ.(ABSTRACT TRUNCATED AT 400 WORDS)

Antigenicity studies of mofezolac (N-22) were examined in mice and guinea pigs and the following results were obtained. The findings of active systemic anaphylaxis, passive hemagglutination test, 4 hour passive cutaneous anaphylaxis (4-hr PCA) and 8-day PCA in guinea pigs revealed that N-22 possessed neither immunogenic nor eliciting potentiality. However, N-22 was shown to be eliciting antigenicity in mice when given N-22-ovalbumin conjugate plus Freund's complete adjuvant (FCA) as immunogen.

A suspension of washed human erythrocytes (2%) in PBS was mixed with an equal volume of 1 mM glutaraldehyde (GA) and allowed to stand at laboratory temperatures, followed by washing with normal saline. The agglutinability of the erythrocytes toward anti-A, anti-B, anti-M and anti-N reagents remained unchanged after GA treatment shorter than 20 minutes, and the agglutinability toward anti-C, anti-c, anti-D, anti-E, anti-e, anti-Lea, anti-Leb and anti-P1 did not decrease after treatment for 10 minutes. GA treatment for longer periods of time than the above caused a decrease of the reactivities. The agglutinability toward an anti-H (Ulex europaeus) and other lectins increased after 10 to 30 minutes of GA treatment and decreased after 40 minutes or more of exposure to GA. These results indicate that the immunologic agglutinability of erythrocytes were practically unchanged after a 10 minutes treatment with 1 mM GA (a mild fixation procedure hereafter called "partial fixation"). The properties of the partially fixed erythrocytes were closely similar to those of untreated erythrocytes as regards cell volume, membrane fragility in hypotonic solutions (measured by a modification of the Ribiere method), and membrane fragility under continuous shaking. The resistance of the partially fixed erythrocytes heating up to 50 degrees C was superior to that of the untreated erythrocytes. From these results, the deformability of the partially fixed erythrocytes was concluded to be similar to that of untreated erythrocytes. After storage for 6 months at 4 degrees C in Alsever solution containing adenine and inosine, the shapes of the untreated erythrocytes changed to "spherocytes" or "echinocytes", whereas the partially fixed cells retained the original discocyte shape. Blood grouping laboratory tests were performed with the partially fixed erythrocytes as indicators. Both anti-A and anti-B agglutinins in normal human sera could be detected without difficulty. Presence of irregular agglutinins, such as anti-H and anti-N, in healthy donors' sera could also be detected, as with the freshly obtained erythrocytes. The detection limits of an incomplete anti-D agglutinin were equal to those in the tests with untreated erythrocytes. The partially fixed erythrocyte (stored at 4 degrees C for 2.5 to 3 months) were used as indicators of ABO-blood grouping from blood stains, saliva stains and hairs by the agglutinin-inhibition test, absorption-elution test or mixed agglutination test. The results obtained were practically equal to those of the tests with freshly obtained erythrocytes, indicating the availabilities of the partially fixed erythrocytes.