New live mycobacterial vaccines: the Geneva consensus on essential steps towards clinical development.

As the disease caused by Mycobacterium tuberculosis continues to be a burden, which the world continues to suffer, there is a concerted effort to find new vaccines to combat this problem. Of the various vaccines strategies, one viable option is the development of live mycobacterial vaccines. A meeting with researchers, regulatory bodies, vaccines developers and manufactures was held to consider the challenges and progress, which has been achieved with live mycobacterial vaccines (either modified BCG or attenuated M.

Advancing TB vaccines to Phase I clinical trials in the US: regulatory/manufacturing/licensing issues.

This article discusses the steps required to file an investigational new drug (IND) application to the United States Food and Drug Administration (FDA) and begin a Phase I trial of a new tuberculosis vaccine in the US. Many different groups play a role in bringing a new vaccine to market. But it is the researcher who begins the process by converting an idea into a new vaccine construct.

A novel approach to develop anti-HIV drugs: adapting non-nucleoside anticancer chemotherapeutics.

Some anticancer drugs, but not all, inhibit replication of human immunodeficiency virus (HIV) and thus, exhibit a therapeutic potential. Such drugs, unlike the traditional HIV enzyme inhibitors, could suppress HIV strains that are resistant to inhibitors of viral enzymes, decrease proviral burden in vivo, or reduce reservoirs of infection via killing infected cells. Thus, they may be an effective adjunct therapy or perhaps result in a cure.

Inhibition of HIV replication by immunoliposomal antisense oligonucleotide.

The sequence-specific suppression of HIV-1 replication using CD4 monoclonal-antibody-targeted liposomes, containing Rev antisense phosphorothioate oligonucleotides is described. Liposomes were prepared by encapsulating the 20-mer antisense DNA sequence of the rev HIV-1 regulatory gene, in the form of a phosphorothioate oligonucleotide. Specific targeting was accomplished by conjugating anti-CD4 mouse monoclonal antibody to the surface of the liposomes.

A novel, sensitive radioimmunoprecipitation assay for the detection of antibodies to human immunodeficiency virus-type 2.

A radioimmunoprecipitation assay that provides an alternative to the Western blot assay was developed for characterizing antibodies against human immunodeficiency virus type-2 (HIV-2). The assay is based on radioiodination of antigen using Bolton-Hunter reagent. The antigen consists of a soluble preparation of the NIH-Z (HIV-2) strain of 1000X purified virus spiked with purified recombinant HIV-2 gp105.

Inhibition of HIV-1 replication in H9 cells by nystatin-A compared with other antiviral agents.

Nystatin A was compared in vitro with amphotericin B, AZT, or foscarnet for their respective abilities to inhibit the replication of human immunodeficiency virus type 1 (HIV-1) in H9 cells. HIV-1-infected H9 cells were cultured for 7 days in the presence of each of these drugs, at various concentrations. Reverse transcriptase activity and p24 antigen production were quantitated.

Evaluation of the single radial-immunodiffusion assay for measuring the glycoprotein content of rabies vaccines.

The glycoprotein content of rabies vaccines containing the Pitman-Moore strain of rabies virus was measured by the single radial immunodiffusion assay and correlated with vaccine potency. The variability of this assay was 6.3% for a single vaccine lot tested over a one-year period.

Inactivation of human T-cell lymphotropic virus, type III by heat, chemicals, and irradiation.

Infectivity of human T-cell lymphotropic virus, Type III (HTLV-III) was inactivated by heat more rapidly if in liquid medium than if lyophilized and more rapidly at 60 degrees than 56 degrees C. When HTLV-III was added to factor VIII suspension, then lyophilized and heated at 60 degrees C for 2 hours or longer there was elimination of 1 X 10(6) in vitro infectious units (IVIU) of virus. Much of the viral inactivation appeared to result from lyophilization.

Selective defects in cytomegalovirus- and mitogen-induced lymphocyte proliferation and interferon release in patients with acquired immunodeficiency syndrome.

To examine the defect in cellular immunity in patients with acquired immunodeficiency syndrome (AIDS), we studied in vitro lymphocyte proliferation and interferon (IFN) release in response to cytomegalovirus (CMV) antigen and Concanavalin A mitogen in 40 homosexual men with AIDS, 10 homosexual men with chronic lymphadenopathy syndrome, 7 healthy homosexual men, and 18 healthy heterosexual subjects of either sex. CMV serology by an enzyme-linked immunosorbent assay and viral cultures for CMV were performed. Lymphocytes of patients with AIDS showed impaired CMV-specific release of IFN but normal mitogen-induced IFN release.

Development of subtype-specific and heterosubtypic antibodies to the influenza A virus hemagglutinin after primary infection in children.

Children undergoing primary infection with an H1N1 or H3N2 influenza A virus developed subtype-specific hemagglutination inhibition antibodies and enzyme-linked immunosorbent assay antibodies to purified hemagglutinin (HA) of the infecting virus subtype. They also developed lower titered ELISA antibodies to the noninfecting H1 or H3 HA and to H8 (an avian strain) HA. Thus, after primary infection with an influenza A virus, children develop enzyme-linked immunosorbent assay, but not hemagglutination inhibition, antibodies reactive with heterosubtypic HAs.

Use of the enzyme-linked immunosorbent assay to detect serum antibody responses of volunteers who received attenuated influenza A virus vaccines.

Sera from volunteers who received live influenza A wild-type or ts recombinant virus were tested by hemagglutination inhibition (HI) assay, neuraminidase inhibition (NI) assay, and the enzyme-linked immunosorbent assay (ELISA) to determine which assay system was the most sensitive in detecting an immunological response to infection. The ELISA was performed with inactivated whole virus antigen, and the optical density at each of five serial twofold dilutions of pre- and postimmunization sera was measured. The difference in the amount of ELISA antibody in pre- and postinoculation serum specimens was taken to be proportional to the area between the respective titration curves.

Evaluation and control of vaccines for the National Influenza Immunization Program.

The National Influenza Immunization Program of 1976 offered an ideal opportunity to test the capability of the system in the United States for production and distribution of maximal amounts of inactivated influenza virus vaccine of carefully regulated quality. The four licensed manufacturers were able to produce and distribute greater than 10 million doses of vaccine per week over a 14-week period. Assays showed that the quality of these vaccines was comparable to or exceeded that of vaccines produced in recent years under less stressful circumstances.

Correlation of laboratory studies with clinical responses to A/New Jersey influenza vaccines.

The large, uniformly performed clinical investigations with influenza A/New Jersey vaccines provided an opportunity to correlate results of laboratory tests of vaccine with human reactivity and antibody responses. These vaccines were given to large numbers of subjects under code, and significant differences in immunogenicity and reactivity were observed in unprimed individuals. A single, relatively large dose of intact virus was more immunogenic and reactive than split-virus vaccines in unprimed subjects.

Quantitation of influenza vaccine hemagglutinin by immunoelectrophoresis.

A technique for the quantitation of hemagglutinin (HA) in influenza vaccines by immunoelectrophoresis (IEP) is described. This method gives accurate, and reproducible results and requires approximately 9 h to complete. The HA content of four manufacturers' vaccines correlated with the chick cell agglutination (CCA) values for each manufacturers' vaccine.

Comparative trial of influenza vaccines. I. Immunogenicity of whole virus and split product vaccines in man.

Groups of about 100 persons aged 6 to 88 years were given 1 of 6 commercially prepared whole virus or split-product bivalent (A/England-B/Mass) influenza vaccines and 6 weeks later were given 1 of 5 monovalent (B/Hong Kong) vaccines. Hemagglutination-inhibiting (HI) antibody titers in serum specimens taken 6 and 12 weeks after inoculation were compared to those obtained before immunization. Overall antibody responses in all groups were adequate, yielding HI titers that are associated with relatively good levels of protection from infection.

Comparative trial of influenza vaccines. II. Adverse reactions in children and adults.

Commercially prepared zonally and chromatographically purified bivalent (A/England-B/Mass) and monovalent (B/Hong Kong) inactivated influenza vaccines were given to 438 individuals 6-33 years old. The vaccines had been examined for antigen content by chick cell agglutination (CCA) tests and electron microscopic particle count determinations. Endotoxin and pyrogen content were determined by limulus amebocyte lysate (LAL) and rabbit pyrogenicity assays; and egg-associated protein contamination was estimated by total protein and single radial immunodiffusion assays.

Influenza virus infection in nude mice.

The role of T-cell function in influenza virus infection was studied by aerosol infection of nude mice with an influenza A virus (PR-8 strain). Nude mice died somewhat later than normal mice, and the antibody response of nude mice to the virus was minimal. Furthermore, nude mice did not eliminate the virus, which persisted for relatively long periods (two to three weeks).

Inactivated influenza vaccine efficacy: diminished antigenicity of split-product vaccines in mice.

Groups of 60 to 120 mice were given a single intraperitoneal inoculation of varying dilutions of commercially prepared and licensed bivalent (A/England and B/Mass) and monovalent (A/Aichi or B/Hong Kong) inactivated influenza vaccines, and their antibody responses at 14 days were quantitated by hemagglutination inhibition tests. Split-product vaccines prepared by the treatment of A/England, B/Mass, and B/Hong Kong whole virus with Tween-80 and either tributylphosphate or ether produced significantly lower mean antibody titers than did equivalent whole-virus preparations. The rates of seroconversion (<1:8 to >/=1:8) at the various dilutions tested were also significantly reduced when these split-product vaccines were given.