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Biomedical subjects

K H Eckels

Publications and source records attributed to K H Eckels.

At least 55 records · Page 3Linked to original sources
PubMed1985

The association of enhancing antibodies with seroconversion in humans receiving a dengue-2 live-virus vaccine.

A group of human subjects, some with yellow fever (YF) antibodies, volunteered for testing of a live-attenuated dengue-2 (DEN-2) vaccine. Serum samples taken before DEN-2 vaccination were tested for their ability to enhance infection of human monocytes by DEN-2 virus. A significantly greater proportion of enhancing antibodies (Eab) were found in YF-immune (YFI) individuals (50%) as compared to those with no evidence of flavivirus infection (9.5%). Geometric mean titers of neutralizing and hemagglutination inhibiting antibodies to DEN-2 virus in YFI subjects with Eab were fourfold to seven-fold higher than in the YFI subjects without Eab in prevaccine sera and 10- to 35-fold higher than in non-immune volunteers. Additionally, levels of Eab in prevaccine sera were directly related to antibody titers found in postvaccine sera. The presence of Eab in the serum of a human subject before DEN-2 vaccination was a good predictor of the immune response after vaccination, and may in part be responsible for the higher seroconversion rate in YF immunes (90%) as compared to nonimmunes (61%) receiving this vaccine. This is the first human study to demonstrate that circulating Eab in non-DEN-immune persons is associated with an augmented immune response to DEN virus infection. This finding supports the hypothesis that cross-reactive antibodies against one flavivirus enhance an infection with another closely related flavivirus.

Adjuvants, Immunologic↗
PubMed1984

Human hypersensitivity to a sham vaccine prepared from mosquito-cell culture fluids.

A sham vaccine, prepared with the C6/36 cell line derived from larval Aedes albopictus mosquitoes, was skin tested on 12 volunteers. Although no reactions were observed after prick tests, three immediate reactions did occur after intradermal tests. Subcutaneous administration of the C6/36 sham vaccine was initially performed on the nine subjects who did not demonstrate immediate reactions. Five of these subjects developed delayed reactions at the site of the intradermal test within 12 hr after inoculation. A Prausnitz-Küstner test was performed on the back of one unsensitized subject by use of both unheated and heated sera from three subjects who had immediate skin reactions, three who demonstrated a delayed reaction, and one who had a negative skin response. In the Prausnitz-Küstner test, immediate skin reactions were observed with all seven unheated sera but with none of the heated serum aliquots. The determination that skin reactivity was associated with a heat-labile serum factor suggested the mechanism was IgE-mediated. Under close surveillance, the C6/36 sham vaccine was administered subcutaneously to one volunteer who had previously demonstrated an immediate response. This subject developed an anaphylactic reaction characterized by hives at the site and more distantly from the site of the subcutaneous inoculation. Hypersensitivity to A. albopictus larval antigens is common and precludes the use of C6/36 cell culture as a substrate for viral vaccines.

Cells, Cultured↗
PubMed1984

Dengue virus type 2 vaccine: reactogenicity and immunogenicity in soldiers.

A live dengue virus type 2 (dengue-2) vaccine (PR-159/S-1) was tested for reactogenicity and immunogenicity in a placebo-controlled, double-blind clinical trial involving 98 soldiers. Seroconversion rates based on the development of neutralizing antibody to dengue-2 were 90% in 70 recipients with immunity to yellow fever and 61% in 28 vaccinees without such immunity (P less than .01). Peak titers of neutralizing antibody were three times higher in recipients with antibody to yellow fever virus and persisted in most for at least 18 months. Individuals seroconverting to the vaccine virus more frequently experienced systemic symptoms than those who received placebo (P less than .02). Future users of this dengue-2 vaccine may wish to employ immunization schedules that include preliminary immunization against yellow fever and must be prepared to accept mild vaccine-related symptoms in some recipients.

Adolescent↗
PubMed1984

Antibody response to dengue-2 vaccine measured by two different radioimmunoassay methods.

Two different radioimmunoassays were used to detect virus-specific antibodies in sera from human volunteers inoculated with an attenuated dengue type 2 (DEN-2) vaccine (PR-159/S-1). An indirect radioimmunoassay required purified DEN-2 virions for optimal reactivity but was 10 to 500 times more sensitive than neutralization or hemagglutination inhibition tests. An antibody capture radioimmunoassay was able to utilize crude antigens from either DEN-infected mouse brains or Aedes albopictus cell culture supernatants. When the two radioimmunoassay techniques were compared, the indirect method appeared to be the best assay for immunoglobulin G (IgG), whereas the antibody capture method was more sensitive for IgM detection. Selected human sera were examined for IgG, IgM, and IgA responses by using both techniques at various intervals after immunization. Although there were differences in magnitude, yellow fever immune as well as flavivirus nonimmune volunteers responded to DEN-2 vaccination by demonstrating IgG, IgM, and IgA antibody responses. In the nonimmune group, the most prevalent immunoglobulin detected was IgM, whereas in the yellow fever immune group, the predominant post-DEN-2 vaccine immunoglobulin was IgG. The preponderance of DEN-2-specific neutralizing antibodies were associated with either IgM or IgG according to the immune status of the volunteer. All classes of immunoglobulins attained maximum levels between 21 and 60 days postvaccination. In the majority of volunteers, IgM responses were relatively transient and could not be detected 6 months after immunization, whereas IgG and IgA antibodies were still detectable after this period.

Antibodies, Viral↗
PubMed1984

Selection of attenuated dengue 4 viruses by serial passage in primary kidney cells. IV. Characterization of a vaccine candidate in fetal rhesus lung cells.

A strain of primary dog kidney (PDK)-passaged dengue (DEN) 4 (H-241) virus cloned by terminal dilution (PDK 35-TD3) was propagated in large volumes in fetal rhesus lung (FRhL) cells to produce a candidate vaccine for evaluation in man. Production seed (FRhL p2) and candidate vaccine (FRhL p3) were subjected to rigorous safety tests to exclude contaminating microbial agents. There was no significant monkey neurovirulence of parental or PDK-passaged DEN-4 virus or of control fluid cultures. FRhL-passaged viruses retained the phenotypic characteristics: small (occasional medium) plaque; temperature sensitivity at 38.5 degrees C; and absence of plaque formation in African green monkey kidney cells, cytopathic effect in LLC-MK2 cells, and viral growth in human monocytes. FRhL p2 virus displayed low virulence for monkeys; only one of four animals was viremic and three of four developed low-titered antibody. FRhL p3 virus produced viremia in three monkeys and moderate to high hemagglutination-inhibition and neutralizing antibody titers in all animals. Virus at both passages in FRhL exhibited reduced neurovirulence in suckling mice as compared to parental DEN-4. Because of its safety and desirable monkey virulence attributes PDK 35-TD3 FRhL p3 is recommended for human phase I trial.

Animals↗
PubMed1984

Selection of attenuated dengue 4 viruses by serial passage in primary kidney cells. V. Human response to immunization with a candidate vaccine prepared in fetal rhesus lung cells.

A dengue 4 (strain H241, PDK35-TD3 FRhL p3) vaccine attenuated by passage in primary dog kidney cells followed by passage and final vaccine preparation in DBS-FRhL-2 cells was tested in five yellow fever-immune volunteers. Only two volunteers seroconverted by producing hemagglutination-inhibiting and neutralizing antibodies. Mild illness, compatible with dengue infection was found only in the individuals who later developed antibodies. Both volunteers developed a rash by the 8th day following vaccination, coinciding with a slight elevation in temperature and leukopenia. Additionally, several serum enzymes were elevated during the observation period. Dengue 4 virus was isolated from the blood of the two infected volunteers starting as early as day 5 post vaccination. During the viremic period, which lasted 5 days, phenotypically-changed virus was recovered, indicating genetic instability of the vaccine virus. The clinical disease and immune response in the two infected individuals was probably related to replication of the variant virus. Further testing of this vaccine in its present form is not indicated.

Adult↗
PubMed1983

Dengue 2 vaccine: dose response in volunteers in relation to yellow fever immune status.

A live dengue 2 vaccine was tested in 38 volunteers in an evaluation of the safety, infectivity, and immunogenicity of doses of 10(1.8)-10(5.5) plaque-forming units. Twenty yellow fever-immune and 18 yellow fever-nonimmune individuals received 0.5 ml of vaccine sc. Immunization was dose related in yellow fever-immune volunteers, with a 50% immunizing dose of 10(3.3) plaque-forming units. In the group not immune to yellow fever, some but not all recipients of each vaccine dilution were immunized, and no 50% immunizing dose could be estimated. Volunteers immune to yellow fever developed adequate titers of neutralizing antibody to dengue 2 virus and maintained them for at least three years; those not immune to yellow fever developed lower antibody titers that disappeared within six months in half of the cases. More than 40 isolates of dengue 2 virus from 12 volunteers retained the in vitro growth characteristics of the vaccine virus; this result affirmed the genetic stability of the virus. Common clinical signs in immunized individuals were leukopenia (55%), macular rash (15%), and fever (10%).

Adolescent↗
PubMed1983

Temperature-sensitive events during the replication of the attenuated S-1 clone of dengue type 2 virus.

Temperature-sensitive events occurring during the replication of the attenuated S-1 clone of dengue type 2 virus were examined. The S-1 clone was more thermolabile than the parent virus at the nonpermissive temperature of 38.5 degrees C. Adsorption experiments in fetal rhesus monkey lung cells revealed an inefficient adsorption of S-1 at 38.5 degrees C compared with the parent virus, suggesting an alteration in a thermolabile virion protein important in adsorption. The production of S-1 viral RNA and antigen occurred at the nonpermissive temperature, which indicated that early events in the replication cycle of S-1 were not affected. Release of infectious virus at 38.5 degrees C was not impaired; however, lower amounts of infectious virus in infected cells at the nonpermissive temperature indicated that maturation of the S-1 clone was suppressed.

Adsorption↗
PubMed1982

Dengue-2 vaccine: infection of Aedes aegypti mosquitoes by feeding on viremic recipients.

Colonized Aedes aegypti mosquitoes were fed on voluntary recipients of an experimental, live, attenuated, dengue type 2 (PR 159/S-1) vaccine to estimate the frequency of vector infection and the stability of the virus in mosquitoes. Two volunteers were viremic at the time of mosquito feeding, but only two of 114 mosquitoes that took a viremic blood meal became infected with the vaccine virus. Strains of virus recovered from the bodies of the mosquitoes and the volunteer's blood retained the temperature sensitivity and small plaque growth characteristics of the vaccine virus. Dengue viral antigen was not detectable in any of the mosquito heads by direct immunofluorescence and in vitro virus transmission by droplet feeding was not observed. This experiment showed that vector mosquitoes can be infected with vaccine virus by feeding on viremic vaccinees. Furthermore, the virus is sufficiently stable to retain the in vitro growth characteristics associated with the vaccine virus.

Aedes↗
PubMed1982

Dengue-2 vaccine: oral infection, transmission, and lack of evidence for reversion in the mosquito, Aedes aegypti.

The dengue-2 vaccine virus (S-1), and its parent virus (PR-159), were compared for their ability to infect orally, to replicate in, and subsequently to be transmitted by Aedes aegypti mosquitoes. The vaccine virus was markedly less efficient in its ability to infect mosquitoes orally. After ingesting infectious bloodmeals containing 3, 7 to 8.2 log10MID50/ml of the respective viruses, 56% (220/396) of the mosquitoes became orally infected with the parent virus contrasted with 16% (66/397) for the vaccine virus. None of the 16 infected mosquitoes transmitted the vaccine virus, while 14% (3/22) of the mosquitoes transmitted the parent virus. The vaccine virus remained temperature-sensitive (restrictive temperature 39 degrees C) after orally infecting and replicating in Ae. aegypti mosquitoes.

Administration, Oral↗
PubMed1981

Dengue-2 vaccine: virological, immunological, and clinical responses of six yellow fever-immune recipients.

Six male volunteers, previously immunized with yellow fever vaccine, were inoculated subcutaneously with a live, attenuated dengue-2 virus (PR-159/S-1) candidate vaccine. Five recipients developed viremia 8 or 9 days after vaccination, which lasted 1 to 10 days. The onset of viremia was followed by fever in three people, transient leukopenia in four, and an erythematous rash in one. One volunteer developed an oral temperature of 38.8 degrees C with headache, myalgia, fatigue, and photophobia suggestive of mild dengue fever. All five viremic volunteers developed fourfold or greater rises in serum neutralizing antibody. The sixth volunteer, who had a low titer of preexisting dengue-2 neutralizing antibody, had no viremia, no symptoms, and a modest rise in hemagglutination inhibiting antibody. Virus isolates obtained from plasma retained the small-plaque and temperature-sensitive growth characteristics of the vaccine virus in vitro. In this study, the vaccine virus genetically stable and immunogenic and seemed sufficiently attenuated for additional testing in humans.

Adult↗
PubMed1980

Dengue-2 vaccine: preparation from a small-plaque virus clone.

The S-1 clone of dengue type 2 virus was used for the preparation of a live-attenuated vaccine after passage in DBS-FRhL-2 cell culture. The vaccine virus had a relatively higher replicative capacity at superoptimal temperatures than its precursor virus, S-1, passaged in primary green monkey kidney cells (S-1 PGMK). There was also a tendency for the S-1 vaccine virus to exhibit leakiness at increased temperatures. Another in vitro marker, replication in monkey peripheral blood leukocytes, indicated less host restriction for the S-1 vaccine in comparative assays with S-1 PGMK virus. Mouse virulence appeared to remain stable on passage in DBS-FRhL-2 cells, whereas monkey immunogenicity decreased. Cautious trials of the dengue type 2 S-1 vaccine in humans are indicated.

Animals↗
PubMed1980

Dengue-2 vaccine: viremia and immune responses in rhesus monkeys.

Studies were undertaken in Indian rhesus monkeys (Macaca mulatta) to determine the safety, potency, immunogenicity, and mosquito infectivity of a small-plaque, temperature-sensitive variant of dengue type 2 (DEN-2) virus, a vaccine candidate. Fifteen monkeys were inoculated subcutaneously with the vaccine virus, ten receiving 10(3.1) plaque-forming units (PFU) and five receiving 10(4.5) PFU. After primary immunization, viremia was detected in only one monkey, a recipient of the higher dose of vaccine. The recovered virus had the same growth characteristics as the vaccine strain. Aedes aegypti mosquitoes did not become infected when they were allowed to feed on monkeys that received the lower dose of vaccine. As expected, the immunization produced no evidence of illness in any of the animals. A dose response to vaccine was detected; all five of the high-dose recipients developed neutralizing antibodies, whereas only five of ten low-dose recipients did so. In both groups, neutralizing antibody was often transient. Its presence at 30 days did not always correlate with protection from viremia in those animals challenged 4 to 6 months after vaccination with wild-type DEN-2 virus. However, immunized animals developed anamnestic antibody responses after challenge, and none demonstrated adverse effects to infection. Reimmunization of monkeys 4 months after primary immunization led to the production of low-titered but persistent neutralizing antibody which protected the animals from a wild-type virus challenge.

Animals↗
PubMed1977

Virulence and immunogenicity of a temperature-sensitive dengue-2 virus in lower primates.

Clones of dengue-2 virus were tested for virulence by inoculation of rhesus monkeys and chimpanzees. Although primates showed no overt signs of illness, inoculation with the parent virus or a subline of a large-plaque clone resulted in a viremia lasting 1 to 7 days. By these criteria, sublines of a small-plaque clone were significantly less virulent and produced little or no viremia in primate hosts. Although they had a substantially reduced viremia, primates inoculated with the small-plaque sublines showed stimulation of complement-fixing, hemagglutination-inhibiting, and neutralizing antibodies. The protection afforded rhesus monkeys 3 months after inoculation with two of the small-plaque sublines was demonstrated by a lack of viremia and a failure to escalate preexisting antibody levels after challenge with the parent virus. Both the S-1 subline and the parent virus had a limited capacity to produce central nervous system pathology in monkeys inoculated intrathalamically and intrathecally. Evidence thus far accumulated for primates indicates that the S-1 subline of dengue-2 virus has potential value as a candidate vaccine virus.

Animals↗
PubMed1976

Isolation of a temperature-sensitive dengue-2 virus under conditions suitable for vaccine development.

Dengue virus, type 2, in viremic human sera and after passage in cell cultures produces mixtures of small and large plaques when assayed in LLC-MK2 cells. Clones of dengue virus type 2 obtained by plaque selection in primary green monkey kidney cell cultures were tested for temperature sensitivity in vitro and for virulence by intracerebral inoculation of suckling mice. Sublines of a small-plaque clone were found to have lower nonpermissive temperatures than the parent virus by both plaque formation and release of infectious virus into the culture media. Small-plaque sublines were significantly less virulent in suckling mice than was the parent virus. Sublines from a large-plaque clone were not temperature sensitive and closely resembled parent virus mixed-plaque morphology. When small-plaque sublines were serially passaged using undiluted inocula, reversion occurred as evidenced by the appearance of large plaques and return of mouse virulence. Small-plaque virus could be maintained through several serial passages without reversion by using low-input inocula. Desirable passage history as well as temperature-sensitive and attentuation characteristics of the S-1 small-plaque subline make it appear suitable as a vaccine candidate virus.

Animals↗
PubMed1975

Virion and soluble antigens of japanese encephalitis virus.

Japanese encephalitis virions contain a 58 X 10-3-molecular-weight envelope glycoprotein antigen that can be solubilized with sodium lauryl sulfate and separated from other virion structural polypeptides and viral ribonucleic acid by gel filtration chromatography. The 58 X 10-3-molecular-weight envelope protein is the major antigen responsible for cross-reactivity of the virion in complement fixation tests with other closely related arboviruses. A naturally occurring soluble complement-fixing antigen is found in Japanese encephalitis mouse brain preparations after removal of particulate antigens. After partial purification by gel filtration and isoelectric focusing, the 53 X 10-3-molecular weight soluble complement-fixing antigen is more type specific than the Japanese encephalitis envelope antigen in complement fixation tests. Further, the Japanese encephalitis soluble complement-fixing antigen is stable to treatment with sodium lauryl sulfate and 2-mercaptoethanol, whereas virion complement-fixing antigens are unstable after this treatment.

Amino Acids↗
PubMed1970

Chikungunya virus vaccine prepared by Tween-ether extraction.

Chikungunya virus vaccines prepared by Tween 80 and ether inactivation of virus grown in green monkey kidney cell cultures were shown to be as immunogenic as comparable Formalin-inactivated vaccines. Both types of vaccine stimulated hemagglutination-inhibiting, complement-fixing, and neutralizing antibody and afforded protection to mice against a live virus challenge. It was shown after Tween-ether treatment of chikungunya virus that the infectivity of the virus was lost and the hemagglutinin titer was increased. By characterization of the resultant hemagglutinin by sucrose and cesium chloride density gradient centrifugation, it was found that the extracted particle was smaller in size and greater in density than the parent virus particle. Removal of lipid may account for the alterations in the physical characteristics of the infectious virus particle. Conditions for treatment of chikungunya virus with Tween and ether were found that preserved high titers of hemagglutinin as well as the immunogenicity of the virus preparations.

Animals↗