BHK-21 RABIES VACCINE BHK-21 RABIES VACCINE – A CANDIDATE VACCINE – A CANDIDATE VACCINE

FOR HUMANSFOR HUMANS

Dusan Lalosevic, MD, Ph.D.*Dusan Lalosevic, MD, Ph.D.*

Pasteur Institute in Novi Sad, Serbia

*Present address: Faculty of Medicine and Clinical Center in Novi Sad, Serbia

Country ReportCountry ReportIn the North community In the North community Subotica, a trial of oral Subotica, a trial of oral vaccination started by delta-vaccination started by delta-planes sponsored by planes sponsored by VIRBACVIRBAC

BHK-21 C 13 cell culture is well known substrate BHK-21 C 13 cell culture is well known substrate

for animal rabies vaccine productionfor animal rabies vaccine production

BHK-21 C 13 cells are much BHK-21 C 13 cells are much better producers of rabies better producers of rabies virus than HDC or VERO virus than HDC or VERO cells…cells…

Baer G.M. Improvements in Rabies Post-Exposure Treatment, Dubrovnik, Yugoslavia, 1984, pp. 15-24:

Only BHK-21 rabies Only BHK-21 rabies vaccine induced vaccine induced

interferon (in mice and interferon (in mice and monkeys), HDC did not!monkeys), HDC did not!

Vaccine. 2006 Mar 20;  Immunogenicity and efficacy of an in-house in-house developed cell-culture derived veterinarian rabies vaccine.

Kallel H, Diouani MF, Loukil H, Trabelsi K, Snoussi MA, Majoul S, Rourou S, Dellagi K.

Unite de Developpement de Vaccins Virologiques, Institut Pasteur, 13 Place Pasteur, BP 74, 1002 Belvedere, Tunis, Tunisia.

The efficiency of an inactivated tissue culture rabies vaccine produced on BHK-21 cells, according to an in-house developed process, was evaluated and compared to a commercial cell-tissue culture vaccine (Rabisin). Fifteen experimental dogs from local common breed were duly conditioned during a quarantine period, then vaccinated via the subcutaneous route with 1ml of either the tissue culture vaccine developed in-house or the commercial vaccine Rabisin. The immune response of each dog was monitored for 162 days. Serum-neutralizing antibodies titers to rabies virus were determined by the rapid fluorescent focus inhibition test (RFFIT) which confirmed the strong response of dogs to both vaccines except one dog in the Rabisin group...

… … and rabies vaccine production is and rabies vaccine production is possible in local institutes…possible in local institutes…

… but up today only for animals.

If a BHK vaccine is good for animals for many years of experience, is this vaccine good for humans too?

It’s a question of the WHO and we have the first answer from late professor Sureau…

Nero

Sureau P. Recent Data on Human Immunization Agains Rabies. Symposium on Rabies, Novi Sad, 17-19th April, 1991

At that time, professor Sureau wrote very carefully:

Yes, we regularly induced tumor on the inoculation site of suspension of BHK – 21

cell culture, but…

… … tumor is solitary, enormous, and never produced tumor is solitary, enormous, and never produced metastases.metastases.

Tumor cells are very similar as BHK 21 C 13 Tumor cells are very similar as BHK 21 C 13 in vitroin vitro,,

Summary of tumorigenicity of BHK-Summary of tumorigenicity of BHK-21 C 13 cell culture21 C 13 cell culture

• This tumor have not the first and most important This tumor have not the first and most important criteria for malignancy – metastases formation!criteria for malignancy – metastases formation!

• BHK-21 cells are tumorigenic only for hamsters BHK-21 cells are tumorigenic only for hamsters (except nude mice)!(except nude mice)!

• Only whole live cells are tumorigenic, not DNA, or Only whole live cells are tumorigenic, not DNA, or cell extracts – It’s a transplantation of cells!cell extracts – It’s a transplantation of cells!

• Conclusion: BHK tumor is Conclusion: BHK tumor is in vivoin vivo culture of BHK-21 culture of BHK-21 C 13 cells!C 13 cells!

• Hypothesis: Immunologicaly, hamsters do not Hypothesis: Immunologicaly, hamsters do not recognize BHK-21 cells as tumorigenic! Becouse of, recognize BHK-21 cells as tumorigenic! Becouse of, transplant growing to enormously tumor masses.transplant growing to enormously tumor masses.

DNA contamination?DNA contamination?

• Perrin P, Morgeaux S. Inactivation of Inactivation of DNA by beta-propiolactoneDNA by beta-propiolactone. Biologicals 1995, 23, 207-211.

• ‘‘Product may be considered safeProduct may be considered safe on the basis of … the possible inactivation of any biological activity of contaminating DNA during processing…’ WHO TRS 878, Geneva, 1998, p. 26.

We accepted their We accepted their opinions and opinions and started with “en-started with “en-house” rabies house” rabies vaccine production vaccine production in Pasteur Institute in Pasteur Institute in Novi Sad, Serbia, in Novi Sad, Serbia, for human use, for human use, namednamed

YU BHK Rabivak.YU BHK Rabivak.

Vaccine was attested Vaccine was attested in the National in the National Center for Center for Epidemiology Epidemiology ”Johan Bela” in ”Johan Bela” in Budapest, a WHO Budapest, a WHO accredited accredited Laboratory for Laboratory for vaccine control, vaccine control, under supervision of under supervision of Imre Lontai, MD, Imre Lontai, MD, Ph.D., Head of Ph.D., Head of Rabies Laboratory in Rabies Laboratory in 2001 year, and also 2001 year, and also in…in…

……Russian Academy Russian Academy of Medical Science of Medical Science Institute named by Institute named by M.P. Chumakhov, M.P. Chumakhov,

under supervision of under supervision of professor V.P. professor V.P.

Grachev, and the Grachev, and the result of vaccine result of vaccine potency was over potency was over

2,5 IU/ml2,5 IU/mlThe first lot of vaccine was

re-attested after 5 years storage at +4 and had 2.6

IU/ml

2.6 2.6 IU/mlIU/ml

5 th July 20065 th July 2006

Clinicaly examination on volunteersClinicaly examination on volunteers•Research staff of Pasteur Institute in Novi Sad (mainly are microbiologists),

•External collaborators included an expert in vaccine control and few veterinarians,

•Hunters

All received:

•Three doses of vaccine in days 0, 7, 21;

•99/100 had antibody titre over 0,5 IU by RFFIT after 30 days;

•1/100 received one additional dose and than have titre over 0,5 IU.

Petricciani J.C.Petricciani J.C.

• What will happened if BHK-21 cell culture rabies vaccine is used on humans?

• ““If such vaccines, which cost as little If such vaccines, which cost as little as 0.50 dollars per dose in USA, as 0.50 dollars per dose in USA, could be used for human beings, the could be used for human beings, the result would be an enormous result would be an enormous reduction in (rabies) morbidity…” reduction in (rabies) morbidity…” Petricciani J.C. Ongoing tragedy of rabies. Lancet Vol 342, October 30, 1993, p. 1067.

•Fourteen years are not enough that responsible persons accepted this

proofs, but…

•At this moment, I believe that time for vaccine production on BHK-21 cells for human use, not only for rabies, is very

close.

Thank you for attention!Thank you for attention!