RU2144955C1 - Strain of influenza virus a/17/pert/95/29(h1n1) for production of live anti-influenza intranasal vaccine for adult humans - Google Patents

Abstract

FIELD: medicine, virology, vaccines. SUBSTANCE: invention relates to preparing live anti-influenza intranasal vaccine for adults humans and children. Vaccine strain A/17/Pert/95/29(H1N1) is reassortant. Strain is obtained by breeding epidemic virus A/Pert/13/95(H1N1) with cold-adapted temperature-sensitive attenuation donor-virus A/Leningrad/134/ /17/57(H2N2). Strain A/17/Pert/95/29(H1N1) multiplicates actively in developing chicken embryos at optimal temperature 34 C and shows temperature-sensitivity and cold-adaptability. Reassortant inherited from epidemic virus two genes encoding surface proteins (hemagglutinin and neuraminidase) and from attenuation donor - six genes encoding nonglycosylated proteins. By antigenic properties the strain corresponds to the strain of virus A/Pert/13/95. Strain A/17/Pert/95/29(H1N1) does not show reactogenicity for adult humans at intranasal administration and corresponds to indices of immunogenicity. By biological properties and indices of reactogenicity the vaccine strain A/17/Pert/95/29(H1N1) corresponds to vaccine strains imposed by Pharmacopoeia article FS 42-299 VS-89 for live anti-influenza vaccine for intranasal use in adult humans. EFFECT: strain of virus indicated above, valuable properties of vaccine. 1 tbl

Description

 The invention relates to medical virology and can be used in healthcare for the prevention of epidemic influenza among adults with a live intranasal influenza vaccine from strain A / 17 / Perth / 95/29 (H1N1).

 The known vaccine strain A / 17 / Texas / 91/1/3 (H1N1) - the prototype [1] - lost its antigenic relevance and as a result will not be able to cause a protective reaction during an influenza epidemic caused by a strain similar to influenza A / Perth / 13 / 95 (H1N1).

 The problem to which the invention is directed, is to obtain a vaccine strain of the actual antigenic variety based on the epidemic virus A / Perth / 13/95 (H1N1).

 Vaccine strain A / 17 / Perth / 95/29 (H1N1) was obtained by genetic reassortment of the epidemic virus A / Perth / 13/95 (H1N1) with a cold-adapted strain A / Leningrad / 134/17/57 (H2N2) - attenuation donor - s subsequent selection in the presence of antisera to the virus A / Leningrad / 134/17/57 (H2N2). The attenuation donor is a cold-adapted temperature-sensitive strain of the influenza virus A / Leningrad / 134/17/57 (H2N2), approved for the production of harmless live intranasal vaccines for adults [2].

Using the method of restriction analysis of DNA copies of the genes of the vaccine and parent strains, it was found that the vaccine strain A / 17 / Perth / 95/29 (H1N1) inherited 6 genes encoding internal proteins (PB1, PB2, PA, NP, M, NS) from an attenuation donor A / Leningrad / 134/17/57 (H2N2) and 2 genes encoding surface proteins - HA (hemagglutinin) and NA (neuraminidase) - from the epidemic virus A / Perth / 13/95 (H1N1). The hemagglutinin in rtga is identical to the virus A / Perth / 13/95 (H1N1), to which the antiserum is completely neutralized. The strain is temperature-sensitive (the difference in the rates of infectious activity at 34 ^o C and 40 ^o C is 8.0 Ig EID ₅₀ / 0.2 ml) and cold-adapted (the difference in the rates of infectious activity at 34 ^o C and 25 ^o C is 3.5 Ig EID ₅₀ / 0.2 ml).

 Thus, the presented vaccine strain A / 17 / Perth / 95/29 (H1N1) is characterized by a combination of useful features necessary for the vaccine strain: antigenic specificity of the surface proteins of the epidemic virus A / Perth / 13/95 (H1N1), the genome structure optimal for reassortant vaccine strains, temperature sensitivity and cold adaptation, which correlates with attenuation for humans, characteristic of an attenuation donor.

 The morphology of the strain is polymorphic, typical of the influenza virus.

Strain characteristics
Infectious activity during reproduction in developing chicken embryos at 34 ^o C for 48 h - 8.0 Ig EID ₅₀ / 0.2 ml

 Hemagglutinating activity - 1: 256.

 The strain exhibits genetic stability of biological traits after 5 passages on chicken embryos (when using large infectious doses).

 Information about the vaccine strain A / 17 / Perth / 95/29 (H1N1) is given below in the passport.

Strain passport
1. The name of the strain is A / 17 / Perth / 95/29 (H1N1).

 2. Series - Series 1.

3. The method of production and characterization of parent viruses when presenting a recombinant strain: recombination
a) epidemic virus - A / Perth / 13/95 (H1N1);
b) donor of attenuation - A / Leningrad / 134/17/57 (H2N2).

 4. The number of passages - 5 in the process of recombination.

5. Characterization of the strain before lyophilization:
a) the optimal conditions for reproduction - 34 ^o C, 48 hours;
b) hemagglutinating activity - 1: 256;
c) infectious activity - 8.0 Ig EID ₅₀ / 0.2 ml;
d) sensitivity to inhibitors - inhibitor-sensitive;
d) the difference in the indicators of infectious activity at 34 ^o C and 40 ^o C - 8.0 Ig EID ₅₀ / 0.2 ml;
e) the difference in the indicators of infectious activity at 34 ^o C and 25 ^o C - 3.5 Ig EID ₅₀ / 0.2 ml

g) the structure of the recombinant genome:
- genes from an epidemic virus - HA, NA;
- genes from the attenuation donor - PA, PB1, PB2, NP, M, NS.

6. Characterization of the strain after lyophilization:
a) date of lyophilization - July 2, 1997;
b) the volume of material in the ampoule is 1 ml;
c) the number of doses in a series is 3;
d) infectious activity - 7.5 Ig EID ₅₀ / 0.2 ml;
d) hemagglutinating activity - 1: 256.

 7. The recommended dilution for vaccination is 1: 3.

8. Antigenic specificity:
a) hemagglutinin in rtga - identical to virus A / Perth / 13/95 (H1N1), the antiserum to which is completely neutralized;
b) neuraminidase - identical to virus A / Perth / 13/95 (H1N1) according to restriction analysis of DNA copies of genes.

 9. Harmless to mice with subcutaneous and intraperitoneal administration - harmless.

 10. Bacteriological control of lyophilized material: date: June 23, 1997, the result is sterile.

 11. Control for the absence of extraneous viruses - extraneous viruses are absent.

 The strain of influenza virus A / 17 / Perth / 95/29 (H1N1) arereactogenic and immunogenic for the adult population.

Example 1. When vaccinating 82 adults with an intranasal spray RJ-4M recombinant vaccine strain A / 17 / Perth / 95/29 (H1N1) with infectious activity of 7.5 Ig EID ₅₀ / 0.2 ml in a dilution of 1: 2 in a volume of 0: , 5 ml of strong temperature reactions with an increase in temperature over 38.5 ^o C and average reactions with an increase in temperature in the range of 37.6-38.5 ^o C were not observed. Weak temperature reactions up to 37.5 ^o C were observed in three vaccinated (3.6%). The reactogenicity index (the difference in the percentage of mild temperature reactions in vaccinated vaccines and received placebo) was 0%.

In the control group of 50 people who received a placebo, strong temperature reactions with increasing temperature over 38.5 ^o C and average reactions with increasing temperature in the range of 37.6-38.5 ^o C were not observed. Weak temperature reactions up to 37.5 ^o C were observed in two individuals of the control group (4.0%) (table 1).

 Thus, the vaccine strain A / 17 / Perth / 95/29 (H1N1) in terms of reactogenicity meets the requirements of Pharmacopeia article FS 42-299 BC-89 for vaccine strains for live influenza vaccine for intranasal use for adults.

List of references
1. Strain A / 17 / Texas / 91/1/3 (H1N1) for the production of live influenza intranasal vaccine for adults. RF patent 2084524 from 07.20.97. Published in the Bulletin of inventions N 20 from 07.20.97.

 2 Alexandrova G.I., Klimov A.I. Live flu vaccine. - SPb .: Science. - 1994 .-- 151 p.

Claims (1)

 The strain of influenza virus A / 17 / Perth / 95/29 (H1N1) GISK N 104 (State Institute of Standardization and Control named after L.A. Tarasevich), used to obtain a live influenza intranasal vaccine for adults.

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