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Biomedical subjects

S Yoksan

Publications and source records attributed to S Yoksan.

32 records · Page 2Linked to original sources

Dengue virus serotypic identification using suckling mouse and western blot technique.

The suckling mouse which is used in the classical method to detect and propagate dengue viruses was evaluated in conjunction with the western blot and immunoenzymatic methods to detect the infecting strains of dengue viruses. After intracerebral inoculation of patients' sera into the suckling mice for 7 days, the mice were examined for the presence of dengue proteins, even though the mice did not have the neurological symptoms which usually serve as an indicator for the presence of dengue infection in the mouse brain. With a blind study of a set of 12 specimens, the suckling mice could detect the virus with the same frequency as the mosquito system but in shorter time of incubation period. The whole process to identify the type of infection takes 9 days. Another important finding is the demonstration of the virion antigen in the liver. The quantity and quality of viral proteins in liver are comparable to those in the brain suggesting that the virus may replicate in the liver as well as in the brain.

Animals↗

Dengue-4 vaccine: neurovirulence, viraemia and immune responses in rhesus and cynomolgus monkeys.

A dengue 4 (DEN-4, strain 1036-PDK 48) vaccine attenuated by passage in primary dog kidney cells was tested using rhesus (Macaca mulatta) and cynomolgus (M. fascicularis) monkeys to determine its safety, potency, and immunogenicity. 14 rhesus monkeys were divided into 3 groups: group 1, 2 animals given control culture fluid; group 2, 2 animals given DEN-4 parental virus; group 3, 10 animals given DEN-4 vaccine virus. 10 cynomolgus were similarly grouped, but group 3 contained 6 monkeys. No significant neurovirulence was observed with parental or with DEN-4 virus passaged in primary dog kidney (PDK) cells. Both cynomolgus monkeys inoculated with DEN-4 vaccine virus developed minimal (V-1) and mild (V-2) neurovirulence-type lesions in the central nervous system, which were nondestructive in both species. All parental and vaccine viruses produced moderate to high neutralizing antibody titres. Only parental virus produced viraemia, in 2 cynomolgus monkeys. Because of its safety and avirulence in monkeys, PDK 48 is recommended for human trial.

Animals↗

Neurovirulence detection of dengue virus using rhesus and cynomolgus monkeys.

The results of a comparative study of neurovirulence of dengue type 1 virus in two species of Old World monkeys, viz. rhesus monkeys (Macaca mulatta) and cynomolgus monkeys (Macaca fascicularis) are reported. In the present study, parental dengue type 1 (16007) and its vaccine viruses were tested by intrathalamic, intramuscular and intraspinal injections in these two species of monkey. Both species of monkeys inoculated with parental dengue type 1 virus developed neurovirulence-type lesions which were graded as minimal (V-1) and occasionally mild (V-2, in cynomolgus monkeys) in severity. The antibody response to either parental or vaccine virus was slightly less in rhesus monkeys than in cynomolgus inoculated with these strains. This comparative study possibly establishes the cynomolgus monkey as a suitable test model to replace the rhesus monkey for neurovirulence testing of dengue-1 vaccine intended for use in humans.

Animals↗

Application of peroxidase-antiperoxidase (PAP) staining for detection and localization of dengue-2 viral antigen. II. Observations for the antibody enhancement activity in human monocytes.

Peroxidase-antiperoxidase (PAP) staining was applied to measure the antibody enhancement activity in human monocytes. Increasing in number of infected cells can be seen with increasing of staining intensity of the cells by ordinary light microscope. Shifting of the optimum enhancement activity was found in previously tritiated antiserum indicated that for titration of antibody enhancement activity several dilutions of antiserum should be included in each experiment. Validity of the PAP method was made by the comparison of the results with Infectious Center Assay (ICA). With this technique, titration for antibody enhancement for dengue virus infection can be done with non-expensive equipment and can be kept for comparison for months.

Animals↗

Immunization with a live attenuated dengue-2-virus candidate vaccine (16681-PDK 53): clinical, immunological and biological responses in adult volunteers.

A live dengue-2 (DEN-2) candidate vaccine (strain 16681-PDK 53), attenuated by passage in primary dog kidney cells, was tested in ten adult volunteers for evaluation of the safety, infectivity and immunogenicity of a dose of 1.9-2.7 x 10(4) plaque-forming units. Five of the volunteers were nonimmune to either dengue or Japanese encephalitis (JE) viruses; the other five were nonimmune to dengue but immune to JE. After receiving 1.0 ml of the vaccine subcutaneously, all ten volunteers developed neutralizing antibodies to DEN-2 which were maintained for at least one and a half years. None of the subjects developed abnormal signs or symptoms and the results of clinical chemistry investigations were within normal range throughout the 21 days of observation after the immunization. Virus isolated from one viraemic volunteer retained the small-plaque and temperature-sensitive growth characteristics of the vaccine virus in vitro. Further testing of this candidate vaccine in humans is indicated.

Adolescent↗

Neurovirulence effects of dengue-2 viruses on the rhesus (Macaca mulatta) brain and spinal cord.

Vaccines prepared from attenuated virus can cause symptomatic viral infection of the central nervous system. In the present study, dengue-2 parental and its live attenuated viruses were tested by intrathalamic and intraspinal injections in rhesus monkeys. The dengue-2 viruses were found to be only very weakly neurovirulent when injected directly into the brain or spinal cord of rhesus monkeys.

Animals↗

Increased phagocytosis of Plasmodium falciparum-infected erythrocytes with haemoglobin E by peripheral blood monocytes.

Erythrocytes from subjects with homozygous and heterozygous haemoglobin E (HbE) infected with Plasmodium falciparum in vitro were phagocytosed to a greater extent by normal human monocytes than infected erythrocytes from normal subjects. Susceptibility to phagocytosis was maximal when the parasites developed to trophozoite and schizont stages in both normal and patients' erythrocytes. The increased susceptibility of P. falciparum-infected HbE erythrocytes to phagocytosis by monocytes may play a role in protection against malaria.

Cells, Cultured↗

Blood value changes in flavivirus-inoculated Macaca fascicularis monkeys.

Blood values were analysed in eighteen cynomolgus monkeys on pre-and post-neurovirulence testing of dengue-2 and yellow fever vaccine viruses, dengue-2 parental and Japanese encephalitis viruses. Certain changes between blood chemistry, hematology and serology were observed and briefly discussed.

Animals↗

Crossed immunoelectrophoresis for the detection of split products of the third complement component in dengue hemorrhagic fever. II. In vitro activation by dengue viral antigen.

The ability of dengue viral antigen(s) from different sources, and performed antigen-antibody complexes, to activate the third component of human complement (C3) in vitro was demonstrated by using split products of human C3. Dengue type 2 viral antigen(s) from suckling mouse brain and from infected monocytes activated C3 via the alternative pathway, while performed soluble dengue-2 antibody complex activated mainly the classical pathway. The implications of this finding for the pathogenesis of dengue hemorrhagic fever are discussed.

Animals↗

Absence of dengue 2 infection enhancement in human sera containing Japanese encephalitis antibodies.

Sera from 52 young adults resident in a rural area in North Thailand were studied for plaque-reducing neutralizing antibodies against dengue (DEN) viruses types 1-4 and Japanese encephalitis (JE), and for DEN-2 infection-enhancing antibodies using a newly described microtest in the human monocyte cell line, U-937. Infection-enhancing antibody titers in U-937 cells using a simplified micromethod were similar to results obtained by published methods using human peripheral blood leukocytes and a macrotest using U-937 cells. In the sample, there were 23 with antibodies to one or more DEN viruses with or without accompanying JE antibodies; 16 sera demonstrated antibodies only to JE and 13 had no detectable antibodies to any flavivirus. All but two DEN antibody-containing sera enhanced DEN-2 infections in U-937 cells, often to titers of 1:10,000 or greater. By contrast, only one of 16 JE-immune sera enhanced DEN-2 infection in monocytes, and that at a dilution of 1:100. None of the flavivirus-negative sera had DEN-2 enhancing activity. The failure of human anti-JE contrasts with the ability of rabbit anti-JE to enhance DEN-2 infections, but correlates with the absence of recorded instances of dengue shock syndrome in human beings sequentially infected with JE and then a DEN virus. This report seemingly reconciles in vitro and in vivo phenomena, and may provide an opportunity to study mechanisms involved.

Adult↗

Application of peroxidase-antiperoxidase (PAP) staining for detection and localization of dengue-2 antigen. I. In an endogenous peroxidase containing cell systems.

The unlabelled immunoperoxidase, peroxidase-anti-peroxidase (PAP), technique was used to detect dengue type-2 viral antigen in several cell systems including the endogenous peroxidase containing cells. These cells are the mosquito cell line (C6/36), continuous cell line of rhesus monkey kidney (LLC-MK2), human monocyte culture both cell suspension and monolayer, and human peripheral blood leukocytes. All of these specimens gave the same results that dengue-2 viral antigen presented in cytoplasm only and the patterns of marker presentation in positive cells varied depending on the duration after infection. The sensitivity of this method is extremely high since it can detect dengue-2 antigen after its attachment on mosquito cells (15 min) as seen in experiments with mosquito cell line, C6/36. False positive was not observed in all cell systems tested.

Animals↗

The micro-focus reduction neutralization test for determining dengue and Japanese encephalitis neutralizing antibodies in volunteers vaccinated against dengue.

A micro-focus reduction neutralization test (mFRNT) was evaluated as an alternative test to the ordinary plaque reduction neutralization test (PRNT) for the determination of dengue virus and Japanese encephalitis virus neutralizing antibody responses in persons receiving dengue vaccine. The 2 tests were similar in terms of titres and ability to detect seroconversion. Although the neutralizing antibody titres obtained by mFRNT were slightly lower than those given by PRNT, the differences were less than two-fold, indicating than mFRNT was reliable. Reproducibility of mFRNT was confirmed by 10 replicate tests using the same control serum. Therefore, mFRNT may be useful in large-scale investigations of neutralizing antibody levels, for example, in young children forming part of an immunization programme; it can be performed quickly and is economical, requiring only a small volume of sera.

Antibodies, Viral↗