Nada Bogdanovic-Sakran

Early phase II trial of human rotavirus vaccine candidate RV3

A naturally attenuated, human neonatal strain, rotavirus vaccine candidate RV3, was tested in a limited phase II randomized double-blind controlled trial. Doses of 1 ml, containing placebo or 6.5 x 10(5) fluorescent cell forming units (fcfu) of virus in AGMK cells, were given at 3, 5 and 7 months of age. Limited replication in the small intestine is implied by the lack of virus excretion, and by the occurrence of an immune response in only 46% of the infants. However, those who developed an immune response were partially protected against rotavirus disease during the subsequent winter epidemic (protective efficacy 54%), supporting observations of protection induced by natural infection by this strain. Protection appeared to be heterotypic. Further trials are warranted, employing strategies to increase immunogenicity of this human rotavirus candidate vaccine.

Genetic and Antigenic Characterization of Rotavirus Serotype G9 Strains Isolated in Australia between 1997 and 2001

ABSTRACT Rotavirus serotype G9 is recognized as the most widespread of the emerging serotypes, emerging since 1996 as a frequent cause of severe acute gastroenteritis in children from many countries covering all continents of the world. This study characterized serotype G9 strains collected in three widely separated Australian centers from 1997 to 2001. All G9 strains possessed the VP4 P[8] and VP6 subgroup II genes. The overall prevalence of the G9 strains increased in Australia, from 0.6% of the strains found in 1997 to 29% of the strains found in 2001. The prevalence of G9 relative to all other serotypes varied from year to year and with geographic location. In Melbourne (representing east coast urban centers), G9 made up 11 to 26% of all of the strains found from 1999 to 2001. In Perth (representing west coast urban centers), G9 made up less than 2% of the strains found in 1997 to 2000 but increased to 18.6% of the strains found in 2001. In Alice Springs (representing widely dispersed settlements in northern arid regions), G9 made up 0 to 5% of the strains found from 1997 to 2000 and was the dominant strain in 2001, making up 68.9% of all of the strains found. Three distinct antigenic groups based on reaction with neutralizing monoclonal antibodies (N-MAbs) were identified, including a dominant group (63%) that cross-reacted with the serotype G4 N-MAb. Phylogenetic analysis of the VP7-encoding gene from Australian strains, compared with a worldwide collection of G9 strains, showed that the Australian G9 strains made up a genetic group distinct from other serotype G9 strains identified in the United States and Africa. Future epidemiological studies of the occurrence of G9 strains should combine reverse transcription-PCR and typing with G1 to G4 and G9 N-MAbs to determine the extent of G9 and G4 cross-reactions among rotavirus strains, in order to assess the need to incorporate G9 strains into new candidate vaccines.

Maternal antibodies to rotavirus: could they interfere with live rotavirus vaccines in developing countries?

INTRODUCTION Past experience with live oral vaccines including licensed rotavirus vaccines demonstrates a trend towards reduced vaccine efficacy in developing countries compared with developed countries. The reasons behind this disparity are not well understood. Transplacental transfer of maternal antibodies and breast milk ingestion may attenuate vaccine responses in infants in developing countries where rotavirus infections are endemic, and maternal antibody levels are high. We examined the prevalence and level of rotavirus antibody in maternal and cord serum, colostrum and breast milk in a developing country setting. METHODS 100 mother-infant pairs were prospectively recruited from December 2008 to February 2009 at Dr. Sardjito Hospital, Yogyakarta, Indonesia. Maternal and cord sera were collected during delivery. Colostrum and transitional breast milk were collected between day 0-3 and day 7-10 postpartum respectively. Rotavirus-specific IgA and IgG were estimated for all specimens and virus neutralization assays were conducted on a subset of milk specimens. RESULTS All maternal and cord serum samples were positive for rotavirus-specific IgG antibodies with a strong correlation between levels of rotavirus-specific IgG in mothers and levels transferred to infants in cord blood (r=0.86; p=0.001). 78% of colostrum and 67% of transitional breast milk specimens were positive for rotavirus-specific IgA. There was a median 4-fold decrease in rotavirus-specific IgA from colostrum to transitional breast milk. Neutralizing antibodies were present in 56% of colostrum specimens assayed (19/34) and in 41% of transitional milk specimens assayed (14/34). CONCLUSIONS Maternal serum and breast milk antibodies to rotavirus are highly prevalent in a developing country setting. Evaluation of the impact of maternal anti-rotavirus serum and breast milk antibody upon vaccine immunogenicity would help to inform rotavirus vaccination strategies, especially in developing settings.

Rotavirus strain surveillance—An Australian perspective of strains causing disease in hospitalised children from 1997 to 2007

This study documents rotavirus strains causing severe disease in Australian children during the pre-vaccine era. During the period 1997-2007, rotavirus strains from national multi-centre hospital-based surveillance in Australia were analysed for G and P types. G1P[8] was the dominant genotype identified during the 11-year study, with intermittent peaks associated with genotypes G2P[4], G3P[8] and G9P[8]. The results provide baseline information of the G and P genotypes causing disease in Australian children, and highlight the unpredictable changes in genotype incidence that can occur on both a local and national level. To be optimally effective, rotavirus vaccines must prevent disease caused by all common rotavirus genotypes.

Hospital-based surveillance for rotavirus diarrhoea in Phnom Penh, Cambodia, March 2005 through February 2007.

Globally rotavirus is the most common cause of severe gastroenteritis in children. From March 2005 through February 2007, a prospective hospital-based surveillance study was conducted at a national hospital in Phnom Penh, Cambodia, to estimate the burden of rotavirus hospitalizations among children aged <5 years old and to determine strain patterns. Children with diarrhoea underwent standard clinical evaluations. Parents were interviewed for demographic and family information. Faecal specimens were tested for rotavirus by enzyme immunoassay (EIA) and positive specimens were further characterized. Of 2817 hospitalized children with diarrhoea, 56% (n=1278) were positive for rotavirus antigen. The G1P[8] strain was the most common genotype (53%) followed by G2P[4] (10%). The findings suggest a need for improved prevention and control programs for rotavirus diarrhoea in Cambodia.

Rotavirus diarrhoea among children aged less than 5 years at Mahosot Hospital, Vientiane, Lao PDR

Rotavirus is one of the most common causes of severe life-threatening diarrhoea in children leading to hospitalization especially in developing countries. At Mahosot Hospital in Vientiane, Lao PDR, children with diarrhoea underwent standard clinical evaluation and faecal specimen collection to estimate the burden of rotavirus hospitalizations and to determine rotavirus strain patterns among children aged less than 5 years old. From March 2005 to February 2007, a total of 1158 stool specimens were collected from children aged less than 5 years old hospitalized with acute diarrhoea. Rotavirus was identified in 624 (54%) of these patients. The G1P[8] strain was the most common genotype (35%), followed by G9P[8] (25%). These surveillance data suggest that improved prevention and control programs for rotavirus as well as other causes of diarrhoea are needed in Lao PDR.

Epidemiology of Rotavirus Diarrhea in Mongolia and Sri Lanka, March 2005–February 2007

BACKGROUND Rotavirus is the most common cause of severe gastroenteritis among children. We conducted hospital-based surveillance to estimate the burden of hospitalizations for rotavirus among children aged <5 years and to describe strain distribution patterns during the 2-year study period. METHODS Children aged <5 years with diarrhea were prospectively enrolled and evaluated by trained pediatricians at representative hospitals in Mongolia and Sri Lanka. Fecal specimens were tested by rotavirus antigen detection enzyme immunoassay. Specimens that tested positive for rotavirus were further characterized to determine the genotype of strains by reverse-transcriptase polymerase chain reaction. RESULTS From 1 March 2005 through 28 February 2007, a total of 1277 hospitalized children with diarrhea were enrolled in Mongolia, and 1916 were enrolled in Sri Lanka. Of the 1152 children in Mongolia who had samples tested, 458 (40%) had results positive for rotavirus, and in Sri Lanka, 428 (24%) of 1806 children with samples tested had positive results. G3P[8] was the most common genotype among rotavirus strains in Mongolia (68%) and Sri Lanka (15%). CONCLUSIONS Rotavirus causes 40% and 24% of hospitalizations for diarrhea among children in Mongolia and Sri Lanka, respectively. Each study site will continue surveillance of rotavirus, and additional laboratory testing will be performed to provide additional information on the distribution of rotavirus strains by G and P genotype.

Characterisation of a G9P[8] rotavirus strain identified during a gastroenteritis outbreak in Alice Springs, Australia post Rotarix™ vaccine introduction

A large rotavirus gastroenteritis outbreak occurred in the Alice Springs region of the Northern Territory, Australia from the 12th of March until the 11th of July 2007. The outbreak occurred five months after the introduction of the Rotarix™ vaccine. Electropherotype and sequence analysis demonstrated that a single G9P[8] strain was responsible for the outbreak and that the strain remained highly conserved during the outbreak period. The outbreak strain contained amino acid changes in regions of the VP7 and NSP4 genes, with known biological function, when compared to previously characterised G9P[8] strains from Australia and other international locations. The recent vaccine introduction was unlikely to have influenced genotype selection in this setting. Importantly, Rotarix™ vaccine was highly effective against the G9P[8] outbreak strain.

Circulating human group A rotavirus genotypes in Malaysia

This study examined the temporal distribution of rotavirus genotypes in Malaysia. Rotaviruses from children with diarrhea admitted to hospitals in 1996 (n = 93) and 2007 (n = 12) in two different regions of Peninsular (West) Malaysia were analyzed for their G and P genotypes using a hemi‐nested RT‐PCR assay. In the 2007 samples, the dominant strain was G9P[8]. It was identified in 42% of the samples. Different strains all possessing the G1 genotype were identified in the rest of the samples. In contrast, 81% of the samples collected in 1996 were the G1P[8] strain. No strains with G9 genotype were detected in samples collected in 1996. J. Med. Virol. 82:707–711, 2010.

Options for improving effectiveness of rotavirus vaccines in developing countries

ABSTRACT Rotavirus gastroenteritis is a leading global cause of mortality and morbidity in young children due to diarrhea and dehydration. Over 85% of deaths occur in developing countries. In industrialised countries, 2 live oral rotavirus vaccines licensed in 2006 quickly demonstrated high effectiveness, dramatically reducing severe rotavirus gastroenteritis admissions in many settings by more than 90%. In contrast, the same vaccines reduced severe rotavirus gastroenteritis by only 30–60% in developing countries, but have been proven life-saving. Bridging this “efficacy gap” offers the possibility to save many more lives of children under the age of 5. The reduced efficacy of rotavirus vaccines in developing settings may be related to differences in transmission dynamics, as well as host luminal, mucosal and immune factors. This review will examine strategies currently under study to target the issue of reduced efficacy and effectiveness of oral rotavirus vaccines in developing settings.