RU2183672C1 - Strain of influenza virus a/17/new caledonia/99/145 (h1n1) for production of live (anti)-influenza intranasal vaccine for adults - Google Patents

Abstract

FIELD: medicine, virology, vaccines. SUBSTANCE: vaccine strain of influenza virus A/17/New Caledonia/99/145 (H1N1) is an reassortant prepared by crossing an epidemic virus A/New Caledonia/20/99 (H1N1) with cold-adapted temperature-sensitive virus A/Leningrad/134/47/57 (H2N2) as a donor of attenuation that is harmless for adults and its properties provide a possibility to obtain reassortant vaccine strains with guarantee from newly appearing epidemic viruses. The strain A/17/New Caledonia/99/145 (H1N1) multiplies actively in developing chicken embryos at optimal temperature 33-34 C, shows temperature- -sensitivity and cold-adaptation. Reassortant inherited two genes from an epidemic virus encoding surface proteins (hemagglutinine and neuraminidase) and six genes encoding nonglycosidated proteins from an attenuation donor. The strain A/17/New Caledonia/99/145 (H1N1) is an areactogenic for adults in an intranasal route of administration. Thus, vaccine strain A/17/New Caledonia/99/145 by biological properties and indices of reactogenicity corresponds to requirements imposing by Pharmacopoeia article PHA 42-3569-98 to vaccine strains for live (anti)-influenza vaccine for intranasal use for adults. EFFECT: new strain indicated above, valuable medicinal properties. 1 tbl, 1 ex

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 / New Caledonia / 99/145 (H1N1). The well-known vaccine strain A / 17 / Perth / 95/29 (H1N1) -prototype [1] - has lost its antigenic relevance and therefore will not be able to cause a protective reaction during an influenza epidemic caused by a strain similar to the influenza A virus / New Caledonia / 20 / 99 (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 / New Caledonia / 20/99 (H1N1).

 Vaccine strain A / 17 / New Caledonia / 99/145 (H1N1) was obtained by genetic reassortment of the epidemic virus A / New Caledonia / 20/99 (H1N1) with a cold-adapted strain A / Leningrad / 134/17/57 (H2N2) - an attenuation donor - followed by 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 / New Caledonia / 99/145 (H1N1) inherited 6 genes encoding internal proteins (PB1, PB2, PA, NP, M, NS) from the donor attenuation A / Leningrad / 134/17/57 (H2N2) and 2 genes encoding surface proteins - HA (hemagglutinin) and NA (neuraminidase) - from the epidemic virus A / New Caledonia / 20/99 (H1N1). Hemagglutinin in rtga is identical to virus A / New Caledonia / 20/99 (H1N1), to which it is completely neutralized by antiserum. The strain is temperature-sensitive (the difference in the rate of infectious activity at 33 ^° C and 40 ^° C is 6.0 lg EID ₅₀ / 0.2 ml) and cold-adapted (the difference in the rate of infectious activity at 33 ^° C and 25 ^° C is 2.0 lg EID ₅₀ / 0.2 ml).

 Thus, the presented vaccine strain A / 17 / New Caledonia / 99/145 (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 / New Caledonia / 20/99 (H1N1), the genome structure is optimal for reassortant vaccine strains, temperature sensitivity and cold adaptation, which correlates with the attenuation for humans, characteristic of the donation of the atenuation.

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

STRAIN CHARACTERISTIC
Infectious activity during reproduction in developing chicken embryos at 33-34 ^o C for 48 hours - 8.0-9.0 lg EID ₅₀ / 0.2 ml

 Hemagglutinating activity -1: 512-1: 1024.

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

 A sample passport for the vaccine strain A / 17 / New Caledonia / 99/145 (H1N1) is attached.

PASSPORT STRAIN
1 The name of the strain is A / 17 / New Caledoyia / 99/145 (H1N1).

 2. Series - Series 1.

3. The production method is recombination; characteristic of parent viruses:
a) epidemic virus - A / New Caledonia / 20/99 (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 - 33 ^o C, 48 hours;
b) hemagglutinating activity -1: 1024;
c) infectious activity - 8.5 lg EID ₅₀ / 0.2 ml;
d) sensitivity to inhibitors - inhibitor-resistant;
d) the difference in indicators of infectious activity at 33 ^o C and 40 ^o C - 6.0 lg EID ₅₀ / 0.2 ml;
f) the difference in the indicators of infectious activity at 33 ^o C and 25 ^o C - 2.0 lg EID ₅₀ / 0.2 ml;
g) the structure of the recombinant genome:
- genes from an epidemic virus - HA, NA,
- genes from an attenuation donor - PA, PB1, PB2, NP, M, NS
6. Characterization of the strain after lyophilization:
a) date of lyophilization - February 28, 2000;
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 lg EID ₅₀ / 0.2 ml;
d) hemagglutinating activity -1: 256.

 7. Recommended dilution with vaccination -1: 3.

8. Antigenic specificity
a) hemagglutinin - identical to virus A / New Caledonia / 20/99 (H1N1) according to RTGA with rat antiserum;
b) neuraminidase - identical to virus A / New Caledonia / 20/99 (H1N1) according to PCR restriction analysis of DNA copies of genes.

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

 10. Bacteriological control of lyophilized material: date: March 16, 2000, the result is sterile.

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

INFLUENZA VIRUS STRAIN A / 17 / NEW CALEDONIA / 99/145 (H1N1) AREACTOGENIC FOR ADULTS CONTINGENT
Example. When vaccinating 50 adults with an intranasal spray RJ-4M recombinant vaccine strain A / 17 / New Caledonia / 99/145 (H1N1) with an infectious activity of 8.5 lg EID ₅₀ / 0.2 ml in a dilution of 1: 3 in a volume of 0.5 ml of strong temperature reactions with increasing temperature over 38.5 ^o C, medium reactions with increasing temperature in the range of 37.6-38.5 ^o C and weak temperature reactions to 37.5 ^o C were not observed.

In the control group of 42 people who received a placebo, strong temperature reactions with a temperature increase of over 38.5 ^o C and average reactions with a temperature increase 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 people (4.76%) (table).

 Thus, the vaccine strain A / 17 / New Caledonia / 99/145 (H1N1) in terms of reactogenicity meets the requirements of Pharmacopoeia article FS 42-3569-98 for vaccine strains for live influenza vaccine for intranasal use for adults.

LITERATURE
1. Kiseleva I.V., Alexandrova G.I., Rudenko L.G., Klimov A.I. The strain of influenza virus A / 17 / Perth / 95/29 (H1N1) for the production of live influenza intranasal vaccine for adults. RF patent N2144955 from 01.27.00. Publ. BI 2000.- N3.

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

Claims (1)

 The strain of influenza virus A / 17 / New Caledonia / 99/145 (H1N1) GISK 396/7 (State Institute of Standardization and Control named after L. A. Tarasevich), used to obtain live influenza intranasal vaccine for adults.

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