Role of lactadherin anti-idiotypic monoclonal antibodies Ab2β in the treatment of human rotavirus-induced diarrhea in a neonatal mouse model / 中国热带医学

@#Abstract: Objective To investigate the preventive and therapeutic effects of anti-idiotypic monoclonal antibodies (Ab2β) of lactadherin on neonatal mice infected with human rotavirus (HRV), and to analyze the underlying mechanism. Methods Hybridoma technology was used to prepare Ab2β of lactadherin. One hundred and twenty 7-day-old Kunming mice were randomly divided into groups A, B, C and control, each consisting of 30 mice. Groups A, B, and C were all infected with HRV via oral gavage. Group A received no treatment, group B was orally administered lactadherin for 7 days prior to infection, and group C was orally administered lactadherin for 7 days after infection, the control group was orally administered cell culture medium that did not contain the virus. The clinical manifestations (diarrhea, body weight) at different time points after infection of the neonatal mice in each group were observed, and the content of rotavirus antigen in the feces of neonatal mice was detected by enzyme-linked immunosorbent assay (ELISA). After HRV infection for 7 days, immunohistochemical staining was used to examine the expression level of intercellular adhesion molecule-1 (ICAM-1) in mouse small intestinal tissues in each group. Results No diarrhea occurred in the control group at any time point. Groups A, B, and C showed diarrhea symptoms after HRV challenge for 1 day. The degree of diarrhea in groups B and C was lower than that in group A at 2-4 days after HRV challenge, and the difference was statistically significant (P<0.05). The HRV antigen content in the feces of the neonatal mice in groups B and C was lower than that in group A at 1-5 days after HRV challenge, and the difference was statistically significant (P<0.05). There was no significant difference in the degree of diarrhea and HRV antigen content between groups B and C at each time point (P>0.05). There was no significant difference in the body weight of the neonatal mice in each group before infection and 1 day after infection (P>0.05); the weight of neonatal mice in groups B and C was higher than that in group A at 3, 5 and 7 days after HRV challenge, and the difference was statistically significant (P<0.05), and there was no significant difference in body weight between groups B and C at each time point after HRV challenge (P>0.05). The number of ICAM-1 expressing cells in the small intestine of the three groups A, B, and C was higher than that of the control group after HRV challenge for 7 days, and the difference was statistically significant (P<0.05). The cell number and gray value of ICAM-1 expressing cells in groups B and C were lower than those in group A, and the difference was statistically significant (P<0.05). Conclusions Anti-idiotypic monoclonal antibodies (Ab2β) of lactadherin has a good preventive and therapeutic effects on human rotavirus infection in neonatal mice, and can significantly improve diarrhea symptoms and reduce HRV viral load. Its specific mechanism may be related to the inhibition of ICAM-1.

Role of lactadherin anti-idiotypic monoclonal antibodies Ab2β in the treatment of human rotavirus-induced diarrhea in a neonatal mouse model / 中国热带医学

@#Abstract: Objective To investigate the preventive and therapeutic effects of anti-idiotypic monoclonal antibodies (Ab2β) of lactadherin on neonatal mice infected with human rotavirus (HRV), and to analyze the underlying mechanism. Methods Hybridoma technology was used to prepare Ab2β of lactadherin. One hundred and twenty 7-day-old Kunming mice were randomly divided into groups A, B, C and control, each consisting of 30 mice. Groups A, B, and C were all infected with HRV via oral gavage. Group A received no treatment, group B was orally administered lactadherin for 7 days prior to infection, and group C was orally administered lactadherin for 7 days after infection, the control group was orally administered cell culture medium that did not contain the virus. The clinical manifestations (diarrhea, body weight) at different time points after infection of the neonatal mice in each group were observed, and the content of rotavirus antigen in the feces of neonatal mice was detected by enzyme-linked immunosorbent assay (ELISA). After HRV infection for 7 days, immunohistochemical staining was used to examine the expression level of intercellular adhesion molecule-1 (ICAM-1) in mouse small intestinal tissues in each group. Results No diarrhea occurred in the control group at any time point. Groups A, B, and C showed diarrhea symptoms after HRV challenge for 1 day. The degree of diarrhea in groups B and C was lower than that in group A at 2-4 days after HRV challenge, and the difference was statistically significant (P<0.05). The HRV antigen content in the feces of the neonatal mice in groups B and C was lower than that in group A at 1-5 days after HRV challenge, and the difference was statistically significant (P<0.05). There was no significant difference in the degree of diarrhea and HRV antigen content between groups B and C at each time point (P>0.05). There was no significant difference in the body weight of the neonatal mice in each group before infection and 1 day after infection (P>0.05); the weight of neonatal mice in groups B and C was higher than that in group A at 3, 5 and 7 days after HRV challenge, and the difference was statistically significant (P<0.05), and there was no significant difference in body weight between groups B and C at each time point after HRV challenge (P>0.05). The number of ICAM-1 expressing cells in the small intestine of the three groups A, B, and C was higher than that of the control group after HRV challenge for 7 days, and the difference was statistically significant (P<0.05). The cell number and gray value of ICAM-1 expressing cells in groups B and C were lower than those in group A, and the difference was statistically significant (P<0.05). Conclusions Anti-idiotypic monoclonal antibodies (Ab2β) of lactadherin has a good preventive and therapeutic effects on human rotavirus infection in neonatal mice, and can significantly improve diarrhea symptoms and reduce HRV viral load. Its specific mechanism may be related to the inhibition of ICAM-1.

Genetic Diversity of Bat Rotaviruses

Rotaviruses are the most common cause of diarrhea in children and animals. Bats are considered reservoirs of many viruses with zoonotic impact worldwide. Rotaviruses have been detected in bats and many of those strains that have been identified globally share high homology with rotavirus strains identified in animals and humans, demonstrating that roles are being created in interspecies transmission and genetic rearrangement in a large number of occasions, which is producing rotavirus genetic diversity. The current effort to characterize strains of rotavirus in bats would help expand knowledge about the great genetic diversity of rotaviruses and could also suggest a bat origin for several unusual rotavirus strains detected in humans and animals. This is a review of the different strains of rotavirus that have been detected in bats globally, where bats have been identified as a possible zoonotic potential in the transmission of rotavirus to animals and humans; and possible anthropozoonosis events are revealed.

Research Progress of Rotavirus and Rotavirus Vaccine / 昆明医科大学学报

Human rotavirus has been discovered since the early 1970s, which is still a primary pathogen causing infant diarrhea under the age of five. Each year about 600,000 infant died of severe body dehydration caused by rotavirus infection. Since the current rotavirus diarrhea have no specific therapeutic drugs, using vaccine to prevent rotavirus infection is an effective method. Research on a vaccine often involved in a multidisciplinary theory and technology. This article reviewed the epidemic situation of rotavirus,animal model,the body's immune response after infection, developing and using vaccine and other related issues.

Application of fecal rotavirus antigen rapid detection in diagnosis of children patients with diarrhea / 国际检验医学杂志

Objective To observe the application effect of fecal human rotavirus (HRV) antigen rapid detection in the diagnosis of children patients with diarrhea .Methods The stool specimens in 200 children cases of diarrhea from January 2014 to December 2015 were collected for detecting HRV antigen by using the gold standard immunochromatographic method .Results Among 200 cases of diarrhea ,129 cases were stool HRV positive with the positive rate of 64 .5% .Among 129 cases of HRV infection ,the ma‐jority were the children patients aged 6 months to 2 years old .The tools in the children patients with HRV infection were egg flow‐er soup like .Conclusion The fecal HRV antigen rapid detection has an important clinical significance for the early diagnosis and early treatment of HRV diarrhea ,so which is worth being promoted in primary hospitals .

Molecular characterisation of the NSP4 gene of group A human rotavirus G2P[4] strains circulating in São Paulo, Brazil, from 1994 and 2006 to 2010

Group A human rotaviruses (HuRVA) are causative agents of acute gastroenteritis. Six viral structural proteins (VPs) and six nonstructural proteins (NSPs) are produced in RV-infected cells. NSP4 is a diarrhoea-inducing viral enterotoxin and NSP4 gene analysis revealed at least 15 (E1-E15) genotypes. This study analysed the NSP4 genetic diversity of HuRVA G2P[4] strains collected in the state of São Paulo (SP) from 1994 and 2006-2010 using reverse transcription-polymerase chain reaction, sequencing and phylogenetic analysis. Forty (97.6%) G2P[4] strains displayed genotype E2; one strain (2.4%) displayed genotype E1. These results are consistent with the proposed linkage between VP4/VP7 (G2P[4]) and the NSP4 (E2) genotype of HuRVA. NSP4 phylogenetic analysis showed distinct clusters, with grouping of most strains by their genotype and collection year, and most strains from SP were clustered together with strains from other Brazilian states. A deduced amino acid sequence alignment for E2 showed many variations in the C-terminal region, including the VP4-binding domain. Considering the ability of NSP4 to generate host immunity, monitoring NSP4 variations, along with those in the VP4 or VP7 protein, is important for evaluating the circulation and pathogenesis of RV. Finally, the presence of one G2P[4]E1 strain reinforces the idea that new genotype combinations emerge through reassortment and independent segregation.

Etiological study on cases with viral diarrhea in Ningxia during 2011 / 中华流行病学杂志

Objective To investigate the etiological characteristics of human rotavirus (HRV),human calicivirus (HuCV),human astrovirus (HAstV) and human enteral adenovirus (HAdV)in Ningxia province during 2011. Methods Stool specimen was collected from acute diarrhea case of Ningxia during 2011. HRV was detected by ELISA and serotype/genotype identified on those RT-PCR positive specimens. HuCV,HAstV and HAdV were detected by RT-PCR. Results In this study,a total of 690 specimens were detected,with the infection rates of HRV,HuCV,HAstV and HAdV as 2.17%,21.74%,3.19%and 6.52%,respectively. Co-infections were found in 4.20%of all the samples being tested. Among 15 HRV positive cases,serotypes G1,G3 and P[4]were the most predominant strains. Conclusion Children who were under 2 years of age were the majority among patients infected by diarrhea viruses while HuCV was recognized as the main pathogen responsible for the viral diarrhea casses in Ningxia,2011.

Etiological study on cases with viral diarrhea in Ningxia during 2011 / 中华流行病学杂志

Objective To investigate the etiological characteristics of human rotavirus (HRV),human calicivirus (HuCV),human astrovirus (HAstV) and human enteral adenovirus (HAdV)in Ningxia province during 2011. Methods Stool specimen was collected from acute diarrhea case of Ningxia during 2011. HRV was detected by ELISA and serotype/genotype identified on those RT-PCR positive specimens. HuCV,HAstV and HAdV were detected by RT-PCR. Results In this study,a total of 690 specimens were detected,with the infection rates of HRV,HuCV,HAstV and HAdV as 2.17%,21.74%,3.19%and 6.52%,respectively. Co-infections were found in 4.20%of all the samples being tested. Among 15 HRV positive cases,serotypes G1,G3 and P[4]were the most predominant strains. Conclusion Children who were under 2 years of age were the majority among patients infected by diarrhea viruses while HuCV was recognized as the main pathogen responsible for the viral diarrhea casses in Ningxia,2011.

Evidence for Genetic Reassortment between Human Rotaviruses by Full Genome Sequencing of G3P[4] and G2P[4] Strains Co-circulating in India

<i>Rotavirus A</i> causes severe diarrhoea in infants and young children worldwide. Many unusual combinations of G and P genotypes have been observed in rotaviruses circulating in developing countries. Mixed infection of a single individual with more than one strain is a mechanism by which genetic reassortants are formed with unusual G and P combinations. However, few studies have provided direct evidence for the formation of such unusual strains as a result of co-infection of co-circulating strains. Here, we used full-genome sequencing to re-analyze a G3P[4] strain (107E1B) and a G2P[4] strain (116E3D) detected in India in 1993 and showed that 107E1B had virtually an identical nucleotide sequence with 116E3D, except the VP7 gene. Phylogenetic analysis revealed that the 107E1B VP7 gene was of typical human rotavirus origin, with a 99.3% nucleotide sequence identity with another Indian G3 VP7 gene. Thus, this study provided robust evidence for the formation of the G3P[4] strain through genetic reassortment in which a G2P[4] strain with a typical DS-1 genogroup background acquired the VP7 gene from a co-circulating G3 human rotavirus strain. This study established a basis on which to facilitate full genome sequence analysis of an increasing number of G3P[4] strains in China and elsewhere in the world.

Prevalence of group A genotype human rotavirus among children with diarrhoea in Nepal, 2009–2011.

Background: Rotavirus as a causative agent of childhood diarrhea is known to cause serious illness among children less than 5 years of age. This study examined the epidemiology of rotavirus disease burden and diversity of G and P genotypes of rotavirus in Nepal. Methods: Stool samples were tested for rotavirus by Enzyme Immuno Assay and Group A rotaviruses were detected by using both ELISA and RT-PCR in 2718 samples between 2009 and 2011. Results: Rotavirus was more frequently detected among inpatients (28.5%) than outpatients (15.2%). Over the three-year study period, 653 (24.4%) cases were positive for rotavirus by ELISA. Genotyping by RT-PCR was done on 638 samples. The most prevalent genotype was G12P [6] (60.4%). Mixed infections were not uncommon (14% in 2009, 29% in 2010 an 7% in 2011). However, 41 were partially typed and 23 were completely untyped over the study period. Conclusions: This study highlights the rotavirus disease burden and diversity of rotavirus strains circulating in Nepal. Continued sentinel surveillance will provide useful information to policy makers with regard to rotavirus vaccine introduction.

Cloning of Full Genome and Genotyping of a Group A Human Rotavirus / 中国生物工程杂志

By means of genetic cloning and recombinant techniques, full genome cDNA sequences of rotavirus strain TB-Chen were isolated from an infantile hospitalized with acute gastroenteritis. Nucleotide sequences analyses showed that the full genome of strain TB-Chen contains 18613 nucleotides, encoding 5791 amino acids. Genotyping results showed that the strain TB-Chen belongs to genotype G2P[4]/NSp4[A]. This is the first report on a full genome of Group A rotavirus in China, and has important significance for deep understanding structure and functions of rotaviruses and developing rotavirus vaccines.

Sequencing VP4, VP7, NSP1, NSP4 genes of human Rotavirus strain G1P8

Rotavirus is the main cause of acute viral gastroenteritis in children under 5 years old. The study is to sequence nucleotides and amino acids of VP4, VP7, NSP1, and NSP4 genes of 5 passages of human rotavirus strain G1P8. The number of nucleotide mutations of VP4, VP7, NSP4 genes occuring among the passages were 3, 1, and 3, respectively. All these mutations resulted in changes in amino acid composition. No mutation was found in NSP1 gene.

Serotype and Nucleotide Analysis of Human Rotavirus Isolates in Korea

Rotaviruses belong to Reoviridae causes diarrhea in human beings as well as domestic animals. This study was conducted to see what type of human rotaviruses are distributed in Seoul and Kyung-gi province. Twenty two of 81 patients showed rotavirus positive with diagnostic kit and RNA electropherosis. We isolated all of rotaviruses from the patients. Electropherotypes of 22 isolates showed 4:2:3 :2 patters whereas those migration patterns were long type. All of those isolates belonged to group 4. Twenty out of 22 isolates reacted with monoclonal antibodies specific to G1, P1A and subgroup II, whereas rest of them, 4-29 and K-30 reacted with subgroup I specific monoclonal antibody. The nucleotide sequence of an isolate K-21 showed 98~100% and 90~96% homologies with those of Wa and KU strain, respectively.

Effect of Bovine and Human Lactoferrin on MA 104 Cell Infected with Human Rotavirus

It has long been known that lactoferrin prevents human beings from infection of virus. To prove this activity of lactoferrin, we evaluated the activities of different lactoferrins to an isolate human rotavirus K-21. Bovine lactoferrin inhibited infection of K-21 to MA-104 cell at the concentration of 25.9 microM whereas bovine hydrolysed lactoferrin prevented rotavirus infection at 103.8 microM. However human lactoferrin prevented infection of K-21 at the concentration of 217.5 microM. These data suggested that lactoferrin activity may be unaffected by the intestinal digestive enzymes and bovine lactoferrin is more active than human lactoferrin with respect to prevention of rotavirus infection.

Serotype and Nucleotide Analysis of Human Rotavirus Isolates in Korea

Rotaviruses belong to Reoviridae causes diarrhea in human beings as well as domestic animals. This study was conducted to see what type of human rotaviruses are distributed in Seoul and Kyung-gi province. Twenty two of 81 patients showed rotavirus positive with diagnostic kit and RNA electropherosis. We isolated all of rotaviruses from the patients. Electropherotypes of 22 isolates showed 4:2:3 :2 patters whereas those migration patterns were long type. All of those isolates belonged to group 4. Twenty out of 22 isolates reacted with monoclonal antibodies specific to G1, P1A and subgroup II, whereas rest of them, 4-29 and K-30 reacted with subgroup I specific monoclonal antibody. The nucleotide sequence of an isolate K-21 showed 98~100% and 90~96% homologies with those of Wa and KU strain, respectively.

Effect of Bovine and Human Lactoferrin on MA 104 Cell Infected with Human Rotavirus

It has long been known that lactoferrin prevents human beings from infection of virus. To prove this activity of lactoferrin, we evaluated the activities of different lactoferrins to an isolate human rotavirus K-21. Bovine lactoferrin inhibited infection of K-21 to MA-104 cell at the concentration of 25.9 microM whereas bovine hydrolysed lactoferrin prevented rotavirus infection at 103.8 microM. However human lactoferrin prevented infection of K-21 at the concentration of 217.5 microM. These data suggested that lactoferrin activity may be unaffected by the intestinal digestive enzymes and bovine lactoferrin is more active than human lactoferrin with respect to prevention of rotavirus infection.

Molecular Cloning and Nucleotide Sequence of the Gene Encoding Fusion(F) Protein of the Thermostable Newcastle Disease Virus Isolated from a Diseased Pheasant

The gene encoding F protein of CBP-1 strain, a heat-stable Newcastle disease virus (NDV) isolated from the diseased pheasants in Korea, was characterized by reverse transcription-polymerase chain reaction (RT-PCR), nucleotide and amino acid sequences. Virus RNA was prepared from the chorioallatoic fluid infected with NDV CBP-1 virus and cDNA was amplified by RT-PCR, cloned and sequenced to analyze. The PCR was sensitive as to detect the virus titer above 25 hemagglutination unit. 1.7kb (1,707bp) size of the cDNA was amplified and cloned into BamHI site of pVL1393 Baculo transfer vector. The nucleotide sequences for F protein were determined by dye terminator cyclic sequencing using four pairs of primers, and 553 amino acid sequences were predicted. In comparison of the nucleotide sequence of F gene of CBP-1 with those of other NDV strains, the homology revealed 88.8%, 98.5% and 98.7% with Kyojungwon (KJW), Texas GB and Beaudette C strains, respectively. As the deduced 553 amino acid sequences of F protein of CBP-1 were compared with those of other NDV strains, the homology appeared 89.9%, 98.7% and 98.9% with KJW, Texas GB and Beaudette C strains, respectively. The putative protease cleavage site (112-116) was R-R-Q-K-R, indicating that CBP-1 strain is velogenic type. The amino acid sequences include 6 sites of N-asparagine-linked glycosylation and 13 cysteine residues. These data indicate that the genotype of CBP-1 strain is more closely associated with the strains of Texas GB and Beaudette C than KJW strain.

A Study of Relation between Serum Aminotransferase Elevation and Clinical Symptoms from Human Rotavirus Gastroenteritis

This study was taken to measure the serum aminotransferase level and its correlation with clinical symptoms from Human Rotavirus gastroenteritis. This report is based on analysis of 434 patients who were admitted to the Pediatric department at Wonju Christian Hospital because of watery diarrhea and vomiting during 2-year period from July, 1991 to June 1993. The stool specimen on admission was tested for Rotavirus Ag by Rotazyme test. And then serum aminotransferase were checked. The result are as follows: 1) Rotavirus Antigens were detected in 194 cases (44.7%) among total 434 patients with gastroenteritis 2) The number of patients with AST elevation above the normal value in RTZ+group was 126 cases(64.9%) and, in RTZ-group was 65 cases(27.0%). 3) The mean value of AST in RTZ+ group was 41.7U/L and, in RTZ-group was 31.1U/L. The mean value of AST between the two groups were significantly different. 4) The mean value of ALT and GGT in the two groups were not different. 5) The patients with AST elevation above normal level in the RTZ+group was significantly increased in frequency and duration of diarrhea than RTZ+ group with normal AST level.

A study of serum transaminase level and it's correlation with several symptoms in children with HRV gastroenteritis

The retrospective study was taken to study the serum transaminase level and it's correlation with several symptoms in human rotavirus gastroenteritis. 494 children, who admitted to the Department of Pediatrics in Dae Dong Hospital from January 1991 to December 1991 with chief complaints of waterdy diarrhea were included in studies. The 1st stool specimen on admission was tested for rotavirus Ag by ELLSA method. and than serum transaminase were checked. The results are as follows: 1) The peak incidence being between 6 months to 2 years in both group, but higher incidence was noted in Non-HRV group. Males are more common than females by ratio of about 2:1. 2) The major symptoms in order of frequency was diarrhea>dehydration>vomiting>coughing>fever in HRV group, diarrhea>dehydration=vomiting>fever>coughing in Non-HRV group. The incidence of dehydration and coughing in HRV group were higher than in Non-HRV group. 3) AST & ALT elevation above the normal value were 83.2% (213/256), 52.0% (133/256) in HRV group and 45.3% (116/238), 22.3% (57/238) in Non-HRV group. AST & ALT were significantly increased in HRV group than Non-HRV group (AST: p0.05, Non-HRV group: p>0.05). 6) Duration of diarrhea for 4-5 days & 1-3 days were relatively more common in HRV group than Non-HRV group. The frequency were 36.3% (93/256) in HRV group and 34.9% (83/268) in Non-HRV group. The degree of diarrhea were not correlated with serum transaminase level at each group (HRV group: p>0.05, Non-HRV group: p>0.05). 7) No fever or duration of fever for 1-2 days were relatively more common in both group. The frequency were 39.9% (102/256), 37.5% (96/256) in HRV group and 38.2% (91/238), 42.5% (101/238) in Non-HRV group. The degree of fever was not correlated with serum transaminase level in HRV group, but correlated with Non-HRV group (HRV group: p>0.05, Non-HRV group: p<0.05).

Clinical observations on human rotavirus gastroenteritis

No abstract available.