Lower Neutralizing Antibody Titers Research Articles

Picornavirus-specific CD4+ T lymphocytes possessing cytolytic activity confer protection in the absence of prophylactic antibodies

Picornaviruses are a family of positive-strand RNA viruses that are responsible for a variety of devastating human and animal diseases. An attenuated strain of mengovirus (vMC24) is serologically indistinguishable from the lethal murine wild-type mengovirus and encephalomyocarditis virus (EMCV). Immunogen-specific stimulation of vMC24-immune splenocytes in vitro demonstrates preferential activation of CD4+ lymphocytes. vMC24-immune splenocytes adoptively transferred to naive recipients conferred protection against lethal EMCV challenge. Immune splenocytes, expanded in vitro, were > 92% CD4+ T lymphocytes. Interestingly, adoptive transfer of these expanded cells engendered protection against lethal challenge. In vivo depletion of CD4+ T lymphocytes prior to lethal challenge abrogated survival of transfer recipients, confirming that CD4+ T lymphocytes were essential for protection. Subsequent rechallenge of vMC24-immune splenocyte recipients with a greater EMCV dose elicited serum neutralizing antibody titers paralleling the high titers observed in vMC24-immunized mice. Unexpectedly, an augmented humoral response was absent in vMC24-specific CD4+ T-cell recipients after the secondary challenge. Moreover, comparably low serum neutralizing antibody titers failed to protect passive transfer recipients when correspondingly challenged. vMC24-immune splenocytes expanded in vitro (> 94% CD4+) lysed vMC24-infected A20.J target cells. The ability to transfer protection with primed CD4+ T cells, in the absence of primed B lymphocytes or immune sera, is novel for picornaviral infections. Consequently, mechanisms such as CD4+ cytolytic T-lymphocyte activity are implicated in mediating protection.

DSPA alpha — Properties of the plasminogen activators of the vampire bat Desmodus rotundus

Summary The saliva of the vampire bat Desmodus rotundus contains a family of potent plasminogen activators, which were named DSPAs after their original source (Desmodus salivary plasminogen activators). The DSPAs share a high structural homology with t-PA. Four different variants of DSPA have been discovered, called DSPA alpha1, alpha2, beta and gamma, out of which, for its unique thrombolytic properties, DSPA α1 has been selected for further development. It has a high potential to develop into a third generation thrombolytic drug. DSPA α1 is a protein with high thrombolytic potency and efficacy. Unlike all other plasminogen activators currently in use, it is almost completely fibrin specific. Furthermore, its prolonged half-life seems to warrant single bolus administration. Above all, recanalization with DSPA α1 seems to occur more rapidly and more completely and, unlike with other plasminogen activators, is sustained over prolonged time periods. Despite being a non human protein its antigenicity is low and only repeated administrations of the substance in short time intervals may non consistently produce low titers of non neutralizing antibodies. The favorable thrombolytic profile of DSPA α1 warrants studying in future thrombolytic trials.

Protection of cats from infectious peritonitis by vaccination with a recombinant raccoon poxvirus expressing the nucleocapsid gene of feline infectious peritonitis virus.

Feline Infectious Peritonitis Virus (FIPV) is a coronavirus that induces an often fatal, systemic infection in cats. Various vaccines designed to prevent FIPV infection have been shown to exacerbate the disease, probably due to immune enhancement mediated by virus-specific immunoglobulins against the outer envelope (S) protein. An effective vaccine would be one that induces cell-mediated immunity without disease enhancing antibodies. In this report, we describe the use of a recombinant raccoon poxvirus that expresses the gene encoding the nucleocapsid protein of FIPV (rRCNV-FIPV N) as an effective vaccine against FIPV-induced disease. Cats were parenterally or orally vaccinated twice, three weeks apart. Cats were then orally challenged with Feline Enteric Coronavirus (FECV), which induces a subclinical infection that can cause enhancement of subsequent FIPV infection. Three weeks later, cats were orally challenged with FIPV. The FIPV challenge induced a fatal infection in 4/5 (80%) of the controls. On the other hand, all five cats vaccinated subcutaneously with rRCNV-FIPV N showed no signs of disease after challenge with FIPV. Four of the five subcutaneous vaccinates survived an additional FIPV challenge. Vaccination with rRCNV-FIPV N induced serum IgG antibody responses to FIPV nucleocapsid protein, but few, if any, FIPV neutralizing antibodies. In contrast to the controls, protected vaccinates maintained low FIPV serum neutralizing antibody titers after FIPV challenge. This suggests that the protective immune response involves a mechanism other than humoral immunity consisting of FIPV neutralizing antibodies.

Mapping of homologous, amino-terminal neutralizing regions of human T-cell lymphotropic virus type I and II gp46 envelope glycoproteins.

Twelve synthetic peptides containing hydrophilic amino acid sequences of human T-cell lymphotropic virus type I (HTLV-I) envelope glycoprotein were coupled to tetanus toxoid and used to raise epitope-specific antisera in goats and rabbits. Low neutralizing antibody titers (1:10 to 1:20) raised in rabbits to peptides SP-2 (envelope amino acids [aa] 86 to 107), SP-3 (aa 176 to 189), and SP-4A (aa 190 to 209) as well as to combined peptide SP-3/4A (aa 176 to 209) were detected in the vesicular stomatitis virus-HTLV-I pseudotype assay. Higher-titered neutralizing antibody responses to HTLV-I (1:10 to 1:640) were detected with pseudotype and syncytium inhibition assays in four goats immunized with a combined inoculum containing peptides SP-2, SP-3, and SP-4A linked to tetanus toxoid. These neutralizing anti-HTLV-I antibodies were type specific in that they did not inhibit HTLV-II syncytium formation. Neutralizing antibodies in sera from three goats could be absorbed with peptide SP-2 (aa 86 to 107) as well as truncated peptides containing envelope aa 90 to 98, but not with equimolar amounts of peptides lacking envelope aa 90 to 98. To map critical amino acids that contributed to HTLV-I neutralization within aa 88 to 98, peptides in which each amino acid was sequentially replaced by alanine were synthesized. The resulting 11 synthetic peptides with single alanine substitutions were then used to absorb three neutralizing goat antipeptide antisera. Both asparagines at positions 93 and 95 were required for adsorption of neutralizing anti-HTLV-I antibodies from all three sera. Peptide DP-90, containing the homologous region of HTLV-II envelope glycoprotein (aa 82 to 97), elicited antipeptide neutralizing antibodies to HTLV-II in goats that were type specific. In further adsorption experiments, it was determined that amino acid differences between homologous HTLV-I and HTLV-II envelope sequences at HTLV-I aa 95 (N to Q) and 97 (G to L) determined the type specificity of these neutralizing sites. Thus, the amino-terminal regions of HTLV-I and -II gp46 contain homologous, linear, neutralizing determinants that are type specific.

Response of colostrum-deprived cynomolgus monkeys to intragastric challenge exposure with simian rotavirus strain SA11

Summary The infectivity and pathogenic potential of a cell culture-adapted simian rotavirus was evaluated in colostrum-deprived newborn and infant cynomolgus macaques (Macaca fascicularis). Intragastric challenge exposure with the simian rotavirus strain SA11 on postpartum day 2 induced diarrhea in 5 of 5 colostrum-deprived newborn monkeys. Compared with sham-inoculated controls, 3 of the 5 inoculated monkeys also manifested reduced body weight gain during the initial 5 days after challenge exposure. Rotavirus was detected in feces of 3 challenge-exposed monkeys for up to 2 days after inoculation. Evaluation of antibody response after rotavirus inoculation was obscured by high but variable prechallenge-exposure serum titers of rotavirus-specific antibody. Preexisting serum titer of neutralizing antibody in newborn monkeys was not predictive of clinical response to inoculation with rotavirus SA11. Two 90-day-old infant monkeys with low serum neutralizing antibody titer did not have diarrhea, reduced weight gain, or antibody response after oral inoculation with rotavirus SA11. Results of these challenge-exposure studies in newborn cynomolgus monkeys were consistent with a heterologous host-rotavirus model and indicate that neonatal serum antibody of maternal origin may not be associated with resistance to rotavirus-induced disease.

SEROLOGICAL EVIDENCE OF HERPESVIRUS INFECTION IN CAPTIVE ASIAN ELEPHANTS (ELEPHAS MAXIMUS)

In mid 1988 a 3-yr-old Asian elephant (Elephas maximus) from a circus in Switzerland died following generalized manifestation of a herpesvirus infection. In an effort to determine prevalence of infection with the herpesvirus, and due to lack of a corresponding virus isolate, it was decided to evaluate contact animals and elephants from a second herd for antibody to bovine herpesvirus 1 (BHV1) and bovine herpesvirus 2 (BHV2). Of 15 sera tested four displayed low neutralizing antibody titers to BHV2. None of the sera neutralized BHV1. However, as evidenced by protein A-mediated immunoprecipitation of metabolically radio-labeled virus-infected and mock-infected cell antigens, followed by separation of precipitation products in SDS-polyacrylamide gels, the 15 sera precipitated multiple antigens from both viruses. Similar results were obtained when using BHV4 antigens. The extent of reaction was most distinct with respect to BHV2 antigens, less prominent with BHV1 antigens, and least with BHV4 antigens. The respective protein patterns, although less marked, matched well with those obtained with bovine reference sera. Additional evaluation of sera from six elephants from two zoos in the Federal Republic of Germany gave essentially identical results. It was concluded that at least one herpesvirus, immunologically related to BHV2, may be widely distributed among captive Asian elephants, and that this virus apparently does not cause overt disease in the majority of animals.

Spectrum of HIV-1 neutralizing antibodies in a cohort of homosexual men: results of a 6 year prospective study.

Serum samples collected prospectively between 1982 and 1987 from a cohort of homosexual men were analyzed for HIV-1 neutralizing antibodies. Seven seroconverters who became infected between 1982 and 1984 and 12 seroprevalents who were already seropositive in 1982 remained free of AIDS. During the 6 year period, 1 seroconverter and 14 seroprevalents developed AIDS. Healthy seroconverters developed neutralizing antibodies slowly and reached moderate geometric mean titers (GMT) of 1:100 within 3 years of seroconversion. Healthy seroprevalents already had moderate titers in 1982, which increased markedly in 1985-1986 (GMT greater than 1:200) and subsequently returned to the previous level in 1987. Neutralizing antibody titers declined 3 years prior to diagnosis in men who developed AIDS and reached levels significantly lower than those of healthy counterparts 2 years before diagnosis. Analysis of neutralizing antibody activity to several HIV-1 isolates indicated that the lower titers of men who developed AIDS were not due to diminished group-specific reactivity. Thus, high neutralizing antibody titers correlated with better clinical outcome, and low or decreasing neutralizing antibody titer signaled poor prognosis. Although naturally developing neutralizing antibodies may ultimately be inadequate in controlling disease progression, measures able to boost the neutralizing antibody titer of individuals already infected with the virus or to elicit high-titer neutralizing antibodies in individuals at risk may yet prove beneficial in controlling viral spread in vivo or in preventing new infection, presumably in concert with cellular immunity.

Characterization of the antibody response in vaccinated mice protected against Coxsackievirus B3-induced myocarditis.

Adolescent male CD-1 mice can be rendered resistant to coxsackievirus B3 (CVB3m)-induced myocarditis following inoculation as neonates with a single dose of a temperature-sensitive mutant virus (ts1), derived from the prototype parent virus (CVB3m). Previously, anti-CVB3 neutralizing antibodies were not detected in sera of adolescent ts1 vaccinees by a standard plaque-reduction assay (Gauntt et al 1983. Infect. Immun. 39:851). However, a more sensitive cytopathic effects-reduction assay permitted detection of low titers of anti-CVB3m neutralizing antibodies of the IgG class prior to challenge with CVB3m. Following CVB3m challenge, serum anti-CVB3m neutralizing antibody titers of ts1 vaccines declined on days 1-2 post-inoculation (p.i.) then increased over the next 6 days. The neutralizing antibodies were of both the IgG and IgM classes. Normal mice challenged with CVB3 did not produce detectable serum anti-CVB3m neutralizing antibody until day 4 p.i. and by 8 days p.i. the neutralizing antibody was only of the IgM class. Thus, adolescent murine ts1 vaccines mount a secondary antibody response to CVB3m in both neutralizing IgG and IgM, but resistance to CVB3m-induced myocarditis is due to the presence of low levels of anti-CVB3m IgG neutralizing antibody in serum at the time of challenge with CVB3m.

Leads from the MMWR. Rabies postexposure prophylaxis with human diploid cell rabies vaccine: lower neutralizing antibody titers with Wyeth vaccine

Leads from the MMWR. Rabies postexposure prophylaxis with human diploid cell rabies vaccine: lower neutralizing antibody titers with Wyeth vaccine

Moloney virus (M-MuLV) leukemogenesis: virus spread, antibody production and antigenic expression in neonatally virus-inoculated young mice.

(A X C57BL) and (A X C57L)F1 hybrid mice were inoculated neonatally with M-MuLV. Virus spread, antigenic expression and antibody production were followed during the preleukemic period. M-MuLV was first detectable in the spleen and later in the thymus. Virus spread was faster and the level of viremia higher in A X C57L than in A X C57BL mice. Also, A X C57L mice had no or only low titers of virus neutralizing antibodies, whereas A X C57BL mice had high titers. Anti-MCSA antibodies, reacting with the surface of syngeneic M-MuLV-induced lymphoma cells, were present in a minority of the mice, but disappeared ultimately in all mice. The two groups of mice differed with regard to the length of the preleukemic latency period. High virus load and a low level of virus neutralizing and anti-MCSA antibodies were correlated with an earlier onset of leukemia.

Experimenten maedi infektion hos får. 2. Bildningen av antikroppar. Alla fyra får som inokulerats via luftvägarna med 7×106 TCID50 av maedi M88 stammen påvisade komplement bindande (CF) antikroppar inom tre månader efter inokulationer, och en gradvis ökning av CF-titern upptäcktes under det första året. Under det påföljande året har CF antikropptitern trots en ringa fluktuation bibehållits och varierat mellan 64

All 4 sheep inoculated via the respiratory tract with 7×106 TCID50 af maedi M88 strain developed complement fixing (CF) antibodies within 3 months after inoculation, and a gradual rise in CF titers was found during the first year. The antibody titers have been maintained, though with some fluctation, through the following year, and the titers vary from 64 to 256. Virus neutralizing activity against maedi M88 strain was detected in the sera of all intrapulmonarily inoculated sheep within 8 months after inoculation. Titers have been maintained or have slightly increased. The level of titers, ranging from 8 to 256, was clearly different between individual sheep. One of the 4 sheep inoculated intracerebrally with 5×105 TCID50 of maedi M88 strain developed CF antibodies 1 month after inoculation, but no neutralizing antibodies until death 11 months after inoculation. The rest of the intracerebrally inoculated sheep displayed no evidence of CF or neutralizing antibodies within 18 months after inoculation in spite of numerous virus isolations from peripheral blood leukocytes. The absence of antibodies might perhaps be attributed to phenomena such as differences in tropism, provirus state, immunological tolerance and size of inoculum. One sheep hyperimmunized with repeated s.c. and i.v. injections of maedi M88 strain developed high CF antibody titers but lower neutralizing antibody titers. The 2 uninoculated control sheep developed no CF or neutralizing antibodies within 18 months after inoculation.

Adjuvant effects of low doses of a nuclease-resistant derivative of polyinosinic acid . polycytidylic acid on antibody responses of monkeys to inactivated Venezuelan equine encephalomyelitis virus vaccine.

Polyriboinosinic.polyribocytidylic acid [poly(I).poly(C)] stabilized with poly-l-lysine and carboxymethylcellulose [poly(ICLC)] has been previously shown to be a compound with marked adjuvant activity when given in high doses with inactivated Venezuelan equine encephalomyelitis (VEE) virus vaccine. This study investigated the effects of much lower doses of poly(ICLC) on the magnitude and kinetics of the primary and secondary humoral antibody responses of rhesus monkeys to inactivated VEE virus vaccine. Monkeys given a single injection of vaccine developed very low neutralizing antibody titers, whereas those given adjuvant plus vaccine had 30- to 100-fold-higher titers which remained elevated for longer than 6 months. Low doses of poly(ICLC) given with VEE virus vaccine resulted in a profound but transient increase in priming of secondary antibody responses to the antigen. In contrast, the administration of poly-l-lysine and carboxymethylcellulose alone without the poly(I).poly(C) component of the complex had no adjuvant effect on antibody responses of monkeys to VEE virus vaccine. The temporal development of antibody by class (immunoglobulin M-immunoglobulin G) in monkeys given two injections of adjuvant-vaccine was not different from that with vaccine alone. Serial hematological and clinical chemistry determinations on monkeys given single or multiple doses of poly(ICLC) with vaccine were not different from values in monkeys given vaccine alone.