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BACKGROUND Human herpesvirus 2 (HHV-2) have DNA genome with a limited genetic variability and have been classified into two clades. OBJECTIVES To identify and characterise six HHV-2 isolates derived from Brazilian women. METHODS HHV-2 isolates were performed polymerase chain reaction (PCR) and sequencing of 2250 pb of the glycoprotein B (gB) coding regions. FINDINGS Four HHV-2 isolates were classified into clade B, while the remaining two, derived from HIV-1 co-infected women, showed a notable genetic divergence (> 1%). MAIN CONCLUSION The results reveal novel HHV-2 variants. The impact of these novel variants on HHV-2 pathogenesis and HIV/HHV-2 coinfection need to be investigated.
Subject(s)
Humans , Female , Herpes Genitalis/virology , HIV Infections/virology , HIV-1 , Herpesvirus 2, Human/genetics , Genes, Viral/genetics , Phylogeny , Herpes Genitalis/complications , HIV Infections/complications , Polymerase Chain Reaction , Bertholletia , Coinfection/virologyABSTRACT
Objective To predict the B cell line epitopes of human cytomegalovirus glycoprotein (gB)by analyzing its structure and physicochemical properties using bioinformatics approaches .Methods Based on the sequence of the HCMV gB,the probable B cell epitopes are predicted using two online prediction programs and DNAstar software .Meanwhile,the tertiary structure of gB was constructed by homologous modeling with the assistance of SWISS -MODEL server to rule out im-possible B cell epitopes .Results and Conclusion The B cell line epitopes of gB are predicted , which provides a theoreti-cal basis for further verification of gB immunodominant epitopes and screening the source plasma with high HCMV IgG titer .
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Background: Molecular epidemiological studies on circulating strains of CMV in cogenital/perinatal infections have not been done earlier in this region. Objective: To study the glycoprotein B genotypes in babies with symptomatic congenital/perinatal CMV infection and to assess the possible influence of genotype on the outcome of the infection. Methods: Clinical samples (blood and urine) of symptomatic babies are sent to the Virology Department of NCDC, Delhi for the diagnosis of congenital infections. 375 clinical samples of infants (newborn - 6 months old) were included for the study. Serum samples were subjected to ELISA for detection of IgM antibodies against CMV. DNA isolation and amplification of CMV genomic DNA targeting gB gene fragment by nested PCR, was carried out in the samples. The amplified fragment including the cleavage site was subjected to RFLP using restriction enzymes Rsal and Hinf1. They were also verified by sequencing using Big Dye Terminator chemistry. Results: 75 samples out of 375 tested were confirmed positive for CMV infection by serology and PCR. Both RFLP and sequencing of gB gene fragment showed that gB 1, 2 and 3 genotypes were in circulation. gB 3 was the most prevalent genotype in symptomatic infants. Hepatosplenomegaly was the most common feature in gB-3 genotype of CMV. gB2 congenital CMV infection was more commonly associated with long term sequelae.
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Human cytomegalovirus glycoprotein B (gB) represents a target for diagnosis and treatment in view of the role it plays in virus entry and spread. Nevertheless, to our knowledge, rare detection of a gB antigen has been reported in transplant patients and limited information is available about diagnostic gB monoclonal antibodies (mAbs). Our aim was to develop gB mAbs with diagnostic potential. Hydrophilic gB peptides (ST: amino acids 27-40, SH: amino acids 81-94) of favorable immunogenicity were synthesized and used to immunize BALB/c mice. Two mAbs, named ZJU-FH6 and ZJU-FE6, were generated by the hybridoma technique and limited serial dilution and then characterized by indirect ELISA, Western blotting, immunoprecipitation, and immunohistochemical staining. The mAbs displayed high titers of specific binding affinities for the ST and SH synthetic peptides at an mAb dilution of 1:60,000 and 1:240,000, respectively. Western blotting and immunoprecipitation indicated that these mAbs recognized both denatured and native gB of the Towne and AD169 strains. The mAbs, when used as the primary antibody, showed positive staining in cells infected with both Towne and AD169 strains. The mAbs were then tested on patients submitted to allogeneic hematopoietic stem cell transplantation. The gB antigen positivity rates of the patients tested using ZJU-FH6 and ZJU-FE6 were 62.0 and 63.0%, respectively. The gB antigen showed a significant correlation with the level of pp65 antigen in peripheral blood leukocytes. In conclusion, two potential diagnostic gB mAbs were developed and were shown to be capable of recognizing gB in peripheral blood leukocytes in a reliable manner.
Subject(s)
Animals , Humans , Mice , Antibodies, Monoclonal , Antibodies, Viral/immunology , Cytomegalovirus Infections/diagnosis , Cytomegalovirus/immunology , Viral Envelope Proteins/immunology , Antibodies, Monoclonal/immunology , Blotting, Western , Enzyme-Linked Immunosorbent Assay , Hematopoietic Stem Cell Transplantation , Immunohistochemistry , Immunoprecipitation , Mice, Inbred BALB C , Sensitivity and SpecificityABSTRACT
Cytomegalovirus (CMV) is one of the most commonly encountered viral pathogens in newborn infants and is found in 0.3-2.4% of all live births. It has been demonstrated that 40-96% of seropositive mothers shed the virus via their breast milk. Breast milk containing CMV can cause almost one-third of CMV infections occurring in infants. A case of postnatal CMV infection in an extremely premature infant (gestational age 24(+5) weeks, birth weight 750 g) transmitted via breast milk is presented. For neonatal intensive care unit (NICU) management of severe thrombocytopenia, anemia, and sepsis syndrome, the infant received repeated transfusions of platelets; intravenous (IV) immunoglobulins; and gamma-irradiated, filtrated packed red cells and was fed her mother's breast milk since the second week of life. CMV infection was diagnosed with positive CMV immunoglobulin M (IgM) and positive urine CMV culture at the second month of life. Considering the negative CMV IgM and urine CMV culture at birth, postnatally-acquired CMV infection was suspected and confirmed with completely identical nucleotide sequence alignments of the infantile blood isolate and the maternal breast milk isolate. To our knowledge, this is the first case of proven postnatal CMV infection transmitted via breast milk in an extremely premature infant in Korea.
Subject(s)
Humans , Infant , Infant, Newborn , Anemia , Base Sequence , Birth Weight , Breast , Cytomegalovirus , Cytomegalovirus Infections , Immunoglobulin M , Infant, Extremely Premature , Infant, Very Low Birth Weight , Intensive Care, Neonatal , Korea , Live Birth , Milk, Human , Mothers , Parturition , Systemic Inflammatory Response Syndrome , Thrombocytopenia , VirusesABSTRACT
Cytomegalovirus (CMV) is one of the most commonly encountered viral pathogens in newborn infants and is found in 0.3-2.4% of all live births. It has been demonstrated that 40-96% of seropositive mothers shed the virus via their breast milk. Breast milk containing CMV can cause almost one-third of CMV infections occurring in infants. A case of postnatal CMV infection in an extremely premature infant (gestational age 24(+5) weeks, birth weight 750 g) transmitted via breast milk is presented. For neonatal intensive care unit (NICU) management of severe thrombocytopenia, anemia, and sepsis syndrome, the infant received repeated transfusions of platelets; intravenous (IV) immunoglobulins; and gamma-irradiated, filtrated packed red cells and was fed her mother's breast milk since the second week of life. CMV infection was diagnosed with positive CMV immunoglobulin M (IgM) and positive urine CMV culture at the second month of life. Considering the negative CMV IgM and urine CMV culture at birth, postnatally-acquired CMV infection was suspected and confirmed with completely identical nucleotide sequence alignments of the infantile blood isolate and the maternal breast milk isolate. To our knowledge, this is the first case of proven postnatal CMV infection transmitted via breast milk in an extremely premature infant in Korea.
Subject(s)
Humans , Infant , Infant, Newborn , Anemia , Base Sequence , Birth Weight , Breast , Cytomegalovirus , Cytomegalovirus Infections , Immunoglobulin M , Infant, Extremely Premature , Infant, Very Low Birth Weight , Intensive Care, Neonatal , Korea , Live Birth , Milk, Human , Mothers , Parturition , Systemic Inflammatory Response Syndrome , Thrombocytopenia , VirusesABSTRACT
BACKGROUND: Human cytomegalovirus (HCMV) glycoprotein B (gB) is the major envelope glycoprotein, encoded by the UL55 gene. Based on sequence variation in the UL55 gene, HCMV can be classified into four gB genotypes. Previous studies have suggested an association between HCMV gB genotypes and clinical outcome in the immunocompromised hosts. The goal of this study was to determine the distribution of HCMV gB genotypes and the effect of gB genotype in the developement of HCMV diseases in hematopoietic stem cell transplant (HSCT) recipients in Korea. MATERIALS AND METHODS: DNA was extracted from 94 blood specimen of 52 allogeneic HSCT recipients with HCMV infection. HCMV gB genotype was determined using polymerase chain reaction to amplify a region of UL55, followed by restriction fragment length polymorphism (RFLP) analysis based on RsaI and HinfI digestion. RESULTS: The distribution of gB types were as follows: gB1, 73.1% (38/52) of patients; gB2, 13.5% (7/52); gB3, 1.9% (1/52) and mixed infection (gB1 and gB2), 9.6% (5/52). While gB4 was not detected, a new genotype (described as gB7 by Trincado et al, 2000) was identified on the basis of their RFLP pattern. During average 708 days' follow up period, HCMV diseases developed in 5 patients. All of them had gB1 genotype. There was no statistically significant association between the incidence of HCMV diseases and the gB genotypes. Re-infection with gB1 strain was detected in one patient who had been previously infected with gB2. This episode was associated with fever, elevated liver enzyme and positive antigenemia. CONCLUSION: HCMV gB1 was the dominant genotype and no gB4 was detected in allogeneic HSCT recipients in Korea, which is an unique pattern compared with the previous reports. Although we can not find significant association between the HCMV diseases and the gB genotypes, genotyping of HCMV will serve in the study of pathogenesis and transmission of this virus in transplant patients. Further study is underway with large study population.
Subject(s)
Humans , Coinfection , Cytomegalovirus , Digestion , DNA , Fever , Follow-Up Studies , Genotype , Glycoproteins , Hematopoietic Stem Cells , Immunocompromised Host , Incidence , Korea , Liver , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , TransplantationABSTRACT
BACKGROUND: Human cytomegalovirus (HCMV) glycoprotein B (gB) is the major envelope glycoprotein, encoded by the UL55 gene. Based on sequence variation in the UL55 gene, HCMV can be classified into four gB genotypes. Previous studies have suggested an association between HCMV gB genotypes and clinical outcome in the immunocompromised hosts. The goal of this study was to determine the distribution of HCMV gB genotypes and the effect of gB genotype in the developement of HCMV diseases in hematopoietic stem cell transplant (HSCT) recipients in Korea. MATERIALS AND METHODS: DNA was extracted from 94 blood specimen of 52 allogeneic HSCT recipients with HCMV infection. HCMV gB genotype was determined using polymerase chain reaction to amplify a region of UL55, followed by restriction fragment length polymorphism (RFLP) analysis based on RsaI and HinfI digestion. RESULTS: The distribution of gB types were as follows: gB1, 73.1% (38/52) of patients; gB2, 13.5% (7/52); gB3, 1.9% (1/52) and mixed infection (gB1 and gB2), 9.6% (5/52). While gB4 was not detected, a new genotype (described as gB7 by Trincado et al, 2000) was identified on the basis of their RFLP pattern. During average 708 days' follow up period, HCMV diseases developed in 5 patients. All of them had gB1 genotype. There was no statistically significant association between the incidence of HCMV diseases and the gB genotypes. Re-infection with gB1 strain was detected in one patient who had been previously infected with gB2. This episode was associated with fever, elevated liver enzyme and positive antigenemia. CONCLUSION: HCMV gB1 was the dominant genotype and no gB4 was detected in allogeneic HSCT recipients in Korea, which is an unique pattern compared with the previous reports. Although we can not find significant association between the HCMV diseases and the gB genotypes, genotyping of HCMV will serve in the study of pathogenesis and transmission of this virus in transplant patients. Further study is underway with large study population.
Subject(s)
Humans , Coinfection , Cytomegalovirus , Digestion , DNA , Fever , Follow-Up Studies , Genotype , Glycoproteins , Hematopoietic Stem Cells , Immunocompromised Host , Incidence , Korea , Liver , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , TransplantationABSTRACT
0.05).Conclusion pcDNA3-gB with different doses have not significant effect on the indexes of hematology,hematological biochemistry and pathology in immunized mice.It is initially proved that pcDNA3-gB is safe.