CH640735A5 - METHOD FOR PRODUCING VIRUS ANTIBODIES. - Google Patents

Description

The present invention relates to antibody-producing cell cultures and in particular to the production of antibodies using living cells in vitro.

The production of specific antibodies for antigenetic determinants of viruses is of great importance for both immunotherapy and medical research.

25 monoclonal antibodies for antigenic determinants of viruses were produced in vitro in cultures of virus-infected spleen cells. So far, however, the antibody production of spleen cell foci in the culture decreased after 30-40 days and stopped completely after 90 days.

We have developed new methods for the production of antibodies; in particular, the cultivation of genetically stable cell lines can be carried out indefinitely in cultures and daughter cultures which produce large amounts of antibodies against viruses and their antigenic determinants. The novel cell lines consist of fused cell hybrids of (a) myeloma cells, i.e. malignant cells from primary tumors of the bone marrow, and (b) virus antibody-producing cells, preferably the spleen or lymph node of animals pre-infected with virus. A particularly preferred cell line is a fused cell hybrid of mouse spleen cells and mouse myeloma cells pre-infected with virus. These cell lines can be maintained in a selective culture medium such as the hypoxanthine-aminopterin-thymidine medium for an almost indefinite period of time and continue to generate the antibodies specific for the antigenic determinants of the virus. In order to grow large amounts of antibodies, in the serum and in the ascites fluid of the animal, the cells can also be grown in vivo in a histocompatible animal.

Myeloma cells are exceptional in themselves because they can produce antibodies, although the specificity of these antibodies is still unknown. The particular species of Tie-55 res from which the myeloma and spleen cells are taken is not critical, since the cells of one species coincide with those of another species, e.g. Mouse-rat, rat-human, can be fused. However, we prefer the same animal species as the source for 60 cells producing myeloma and virus antibodies. Excellent results were achieved with somatic cell hybrids of spleen cells producing virus antibodies from a BALB / c mouse previously immunized with virus and myeloma cells from a BALB / c mouse. Particularly preferred myeloma cells are the cells of a MOPC-21 designated as clone (P3 x 63 Ag8) 65, which were described by Kohler et al. in Nature, 256,495-497 (1975).

Fused cell hybrids of BALB / c spleen cells and BALB / c myeloma cells have previously been described in the literature by

3rd

640 735

Kohler et al. in Nature, 256,495-497 (1975) and in Eur. J. Immunol., 6, 511-519 (1976). However, these hybrids were not derived from mouse immunized mice, but from mice immunized with sheep red blood cells. Hybrids of sheep erythrocyte antibody-producing cells and myeloma cells can be easily produced, but the hybrids according to Kohler et al. generated antibodies specific for sheep erythrocytes and have no influence on viruses. Before this invention it was not known whether hybrids could be developed from virus antibody-producing cells and myeloma cells.

A method for producing a cell line of hybrid cells can be seen from the following, the spleen cells of a BALB / c mouse previously immunized with influenza virus being fused with myeloma cells of a BALB / c mouse. Although a special strain of the influenza virus was used in the method, viruses such as rabies, mumps, cowpox, influenza strain 6-94, Simi-an-Virus 40, polio, etc. can also be used.

a) Preparation of spleen cells for fusion

In order to prepare the spleen cells for the fusion, BALB / c mice, which had been infected 2-3 months beforehand with a purified influenza virus [A.PR/8/34 (HONI)] in a traperitoneal injection of 1250 hemagglutinating units, were administered intravenously a booster of 100 homologous hemagglutinating units. The mice were sacrificed 7 days later and a spleen cell suspension as described by Gerhard et al. , Eur. J. Immunol., V. 720-725 (1975). The red blood cells were lysed by incubation for 15 minutes at 4 ° C. in NH4.CI (0.83%). The resulting cell suspension was washed by centrifugation (800 x g) in heat-inactivated calf serum and centrifugation in protein-free medium (RPMI1640, buffered with 7.5 mM HEPES, pH 7.2).

b) Preparation of myeloma cells for the fusion

BALB / c- (P3 x 63 Ag8) myeloma cells made by the

MOPC-21 line descend and the HPRT (E.C.2.4.2.8) are deficient as by Dr. Cesar Milstein in Nature, 256, 495-497 (1975) was grown in Eagle's minimally idiopathic medium (MIM) containing 10% calf serum and 10% horse serum.

c) Production of hybrids

Hybrid production was achieved by mixing 107 BALB / c (P3 x 63 Ag8) myeloma cells with 108 spleen cells from the virus-infected BALB / c mice. The cell mixture was centrifuged at 800 x g and resuspended for fusion in a 50 percent solution (w / v) of polyethylene glycol-1000 (PEG) and diluted in minimally idiopathic medium (MIM) without serum.

According to the Davidson et al. described fusion methods (Somat, Cell Genet., 2,175-176), the polyethylene glycol was diluted with MIM first without serum and then with serum and then five 75 cm falcon flasks with hypoxanthine-aminopterin-thymidine (HAT) selective-5 Inoculates medium. The cultures were incubated at 37 ° C in an atmosphere of 95% air and 5% CO 2 in the incubator and the culture medium was partially replaced by fresh HAT medium (1/2 to 1/3) every 7-10 days.

Ten to fifteen days after incubation of cultures 10 made by fusion of spleen cells from PR8 immunized mice with P3 x 63 Ag8 cells, cell proliferation was observed in a bottle. Any cell proliferation in HAT medium indicates successful hybridization of mouse spleen and mouse myeloma cells. 15 These cells, designated HK-PEG-1, were continuously grown in HAT medium and cloned on microplates (Linbro) by dilution. The remaining 4 flasks, in which no cell proliferation occurred, were destroyed 3 weeks after the fusion. A cell line preparation designated HK-PEG-1 was deposited with the American Type Culture Collection and has the accession number ATCC CL189. (Filing date: June 15, 1977)

d) Karyological analysis

As can be seen from Table 1, the P3 x 63 Ag8 parent cells contained an average of 63 chromosomes and BALB / c spleen cells 40 chromosomes. The 92 chromosomes in the HK-PEG-1 cells thus corresponded approximately to the total number of chromosomes in both parent cells. The karyology of the hybrid clone HK-PEG1 and the subclones was monitored for 4 months without a significant change in the karyotype being observed.

Table 1

35 Number of chromosomes in parental and hybrid cells

Cells Average number of

Chromosomes per cell

63 *

40 92 *

* including 2 metacentric marker chromosomes.

45

e) Analysis of the immunoglobin produced by the hybrid cells

Culture fluids from HK-PEG-1 and P3 x 63Ag8 cells were examined for the presence of immunoglobulin (Ig) 50, which reacts with PR8 in the radioimmunoassay (RIT). P3 x 63 is known to secrete Ag8 IgGl, k. As can be seen from Table 2, P3 x 63 Ag8 culture fluids contained no Ig with anti-PR8 reactivity. In contrast, large amounts of IgG Anti-55 PR8 antibody were produced by HK-PEG-1.

25th

30th

40 P3 x 63 Ag8 BALB / c spleen cells HK-PEG-1

Table 2

Classes of anti-PR8 antibodies produced by HK-PEG-1

Culture liquid in the RIT * CPM examined in the RIT with 125I-labeled antisera tested Anti-F (ab ') 2 anti-IgM anti-IgA anti-IgG

P3 x 63 Ag8 44 ± 24 ** NE *** NE 185

HK-PEG-1 4255 + 158 ** 204 26 8335

* Medium from cultures with similar cell concentration. The analysis was carried out using replication samples from 15

ul (P3 x 63 Ag8) or 7.5 uL (HK-PEG-1) of the culture liquid.

** Average + SF of 6 investigations. Other entries: average of RIT, carried out twice.

*** Ne = Not determined.

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4th

The information in Table 3 indicates that the virus antibodies (anti-PR8) produced by the hybrid cells are specific for the antigen (hemagglutinin) of the PR8. This can be seen from the functional tests in which the antibodies (approximately 40 μg / ml) brought about an inhibition of hemagglutin (HH) but no detectable inhibition of neuraminidase (NH).

Table 3

Activity of the antibodies against Pr-8 virus produced by HK-PEG-1 (in functional tests)

Antibodies from HK-PEG-1 secreted in vitro and concentrated by ammonium sulfate precipitation **.

HH titer * (log 10) 2.98

NH titer * (log 10) <1.00

* Hemagglutination inhibition (HH) and neuraminidase inhibition (NH) against PR8 virus.

** Fifty ml of HK-PEG-1 culture medium was mixed with ammonium sulfate at 4 ° C with 42% (v / v) saturation. Precipitate was dialyzed against PBS and concentrated to 0.6 ml by ultrafiltration. As determined by RIT, this final sample contained 47% of the antiviral antibodies present in the initial volume of the culture medium. This corresponded to a 40-fold concentration of the antiviral antibodies.

f) Clonic origin of the hybrid cells:

The clonic origin of HK-PEG-1 has been checked against 3 independent criteria:

(i) Concentrated Ig obtained from HK-PEG-1 mass culture (see label, Table 3) was foamed isoelectrically, anti-PR8 antibodies with IgGl heavy chain determinants accumulated in a limited pH range.

(ii) The antibody was tested in the RIT for cross-reactivation with respect to various viruses known to be antigenically related to PR8 and found to be specific for the determinant of PR8 hemagglutinin (HA). However, 20-25% of the PR8 pre-infected spleen precursor B cells have this strain-specific anti-HA (PR8) responsiveness. Antibody responsiveness alone is therefore a weak criterion for determining the monoclonality of HK-PEG-1.

(iii) In order to exclude the possibility that antivirus antibody-producing cells represent only a small fraction 25 of the hybrid cell population, the culture fluids derived from 12 clones of the HK-PEG-1 hybrid line were checked for the presence of anti-PR8 antibodies and their specificity checked. As can be seen from Table 4, 11 of the 12 clones raised anti-PR8 antibodies. Furthermore, these antibodies were specific for a determinant of the HA of the PR8 virus.

Table 4

Anti-PR8 antibodies produced by clones of the HK-PEG-1 cultures

Culture fluid from HK-PEG-1 clone no.

Concentration of anti-PR8

HA antibodies *

(Ug / ml)

2nd

3rd

5

6

7 9

10th

11

12

13 14a 14b

2nd

1.2 2.4 3.4 4.7 6.9

4.7

5.8 <0.02

2.8 2.7 2.6

* RIT-based quantitative analysis performed with 125I-Anti-F (ab ') 2.

g) tumor formation by hybrid cells

To determine tumor formation by hybrid cells, adult mice were injected subcutaneously in the abdominal wall with either HK-PEG-1 (I x 107) or P3 x 63 Ag8 (1 x 107) cells.

As can be seen from Table 5, tumors developed at the vaccination site of the mice injected with the HK-PEG-1 (5/5) and the mice injected with the P3 x 63 Ag8 parent cells (3/5) ten to twelve days after the implant. In a second experiment, 4/5 mice developed tumors after vaccination with hybrid cells and 3/5 after vaccination with 60 P3 x 63 Ag8 cells. After intraperitoneal injection of cells (experiment 3, table 5), it was found that tumors grew in abundance in the abdominal cavity. In order to produce ascitic fluid, it was necessary to inject tumor cells into the mice that were completely resuspended in Freund's adjuvant 65. In a fourth experiment, mice injected with Pristan developed clone ascites after intraperitoneal inoculation with HK-PEG-1 cells.

5

Table 5

640 735

Experiment No. Type of vaccination Vaccine culture Ratio of mice,

who developed subcutaneous tumors or ascites:

1 subcutaneously

2 subcutaneously

3 intraperitoneally

4 intraperitoneal **

* No tumor or ascites 30 days after vaccination. ** Mice injected with Pristan.

The results of these studies show that hybrid cells from myeloma and normal spleen cells in mice behave like the malignant parent without suppressing the malignancy.

After the tumor mice at different times

HK-PEG-1 P3 x 63 Ag8 HK-PEG-1 P3 x 63 Ag8 HK-PEG-1 in CFA HK-PEG-1 HK-PEG-1 Kl 6 HK-PEG-1 Kl 7 HK-PEG-1 Kl 12

5/5 3/5 4/5 3/5

3/3

0/3 4/4 4/4 4/4

Sera and ascitic fluids taken from injection were examined using RIT to determine the concentration of anti-PR8 antibodies. The results are shown in Table 6.

Table 6

Anti-PR8 antibodies in sera and ascitic fluids taken from mice injected with HK-PEG-1 cells

Try days by type of estimated

No injection sample concentration of

Antivirus antibody * mg / ml

1

28

serum

1.35

serum

1.1

serum

2.3

2nd

19th

serum

0.9

serum

1.6

Serum**

1.2

Ascites **

0.5

4th

15

Ascites

0.5

Cl 6

Ascites

0.45

Cl 7

Ascites

<0.002

Cl 12

* RIT-based quantitative analysis performed with 125I-anti-F (ab ') 2.

** From the same mouse.

Sera from mice injected subcutaneously with HK-PEG-1 cells (experiments 1 and 2) contained anti-PR8 antibodies in a concentration of 1 to 3 mg / ml. Anti-PR8 antibodies were also found in the ascitic fluid of mice intraperitoneally injected with hybrid cells (Experiment 4), but the concentration of antibodies was 3 to 4 times lower than in the serum.

Electrophoresis of serum from HK-PEG-1 tumor mice showed the presence of 3 main Ig populations: one like that of the parental (P3 x 63 Ag8) IgGl, one characteristic of IgG3 and a third between the other two. To determine which class of immunoglobulins the antivirus antibody was associated with, ascitic fluid from 20 was injected with pristan

55 mice pooled and the immunoglobulins separated therefrom. The separation was achieved in the following way:

The ascitic fluid removed from the mice was pooled and neutralized with phosphate buffered saline (PBS). An equal volume of saturated ammonium sulfate was added at 4 ° C., the precipitated protein was dissolved in PBS, and dialyzed against 0.01 M Tris buffer at pH 8.0. The precipitate which formed overnight during dialysis was redissolved in PBS and again precipitated by dialysis against 0.01 Tris buffer at pH 8.0. The supernatant of the original dialysate was equilibrated on a DE-52 (Whatman) column with 0.01 M Tris buffer, pH 8.0 and with a linear NaCl gradient (0.05 to 0.15 M NaCl in 0, 01 M-Tris, pH 8.0).

60

65

640735

6

After precipitation of the IgG3 by dialysis against 0.01 M Tris buffer, it was found after electrophoresis that the supernatant lacked a component which was present in the ascitic liquid before the fractionation. Chromotography of this material on DE-52 did not result in a complete separation of the other components, however the electrophoresis of the individual fractions showed the presence of two main components: one corresponding to the parental (P3 x 63 Ag8) IgGl and the other like a Hy -brid-Ig, was an intermediate between IgGl and IgG3, which was also observed in the serum of HK-PEG-1 tumor mice (see above).

The concentration of anti-PR8 antibodies was determined in RIT as 125I-anti-F (ab ') 2 and the specific anti-PR8 activity was calculated as a ratio to anti-PR8 (mg / ml) 2 Ig (mg / ml) , the latter calculation is based on measurements of the absorption at 280 nm with an assumed absorption (1% -I cm) for mouse Ig of 14.0. The specific anti-PR8 activity of the precipitate obtained at low salt concentration was almost twice as high as the maximum of the anti-PR8 activity in DE-52 chromatography (0.06 in fraction 21): analysis of the samples in the RIT with 125I-anti-IgGl revealed that about 15% of the anti-PR8 antibodies that were precipitated at low salt concentration showed Gl determinants. In contrast, 125I-anti-F (ab ') 2 was just as effective in the quantitative determination of anti-PR8 antibodies in the different fractions from DE-52 chromatography.

A number of other attempts have been made with rabies virus. BALB / c mice were injected with inactivated vaccine containing rabies virus (ERA strain) and a booster vaccination intravenously one month later was given to the same vaccine. The mice were sacrificed three days after vaccination and hybrid cells were produced by fusion of the spleen cells with P3 x 63 Ag8 myeloma cells as described above. Many of the hybrid cells produced anti-rabies antibodies. If the mice were killed 10 days after the vaccination, the number of hybrid cells producing anti-rabies antibodies was very low. This method was used to obtain the hybridomas for the tests shown in Tables 7, 8,9.

i5 The rabies strains used in the following experiments are known strains. The ERA strain is e.g. in U.S. Patent 3,423,505 and U.S. Patent 3,585,266 and SAD in Can. J. of Microbiology, 6, 606 (1960). The analytical tests used are also known.

Fifty-two hybridomas, which were identified in the RIT (radioimmune test) and produced anti-rabies antibodies, were tested against 3 different strains of rabies virus (ERA, CVS and HEP-Flury strains). In addition, 4 different test methods were used: virus neutralization (VN), cytotoxic test (CT), membrane fluorescence (M) and nucleocapsid fluorescence (NC). The test results are shown in Table 7.

Table 7

Responsiveness to virus strains in different samples

Number of ERA CVS HEP

Hybridomas VN CT M NC VN CT M NC VN CT M NC

7

+

+

+

-

+

+

+

-

+

+

+

-

2nd

+

+

+

-

±

+

+

-

+

+

+

-

2nd

+

+

+

-

+

+

+

-

-

-

-

-

11

+

+

+

-

-

-

-

-

+

+

+

-

2nd

+

+

+

+

-

-

-

+

+

+

+

+

2nd

+

+

+

+

-

-

-

+

-

-

-

+

1

+

+

+

-

-

-

-

-

-

-

-

-

2nd

-

+

+

+

-

-

-

+

+

+

+

+

1

-

-

-

-

-

-

-

-

+

+

+

-

22

-

-

-

+

-

-

-

+

-

-

-

+

As shown in Table 7, antibodies that react with one strain cannot affect another rabies virus. As can be seen from Table 7, 9 out of 52 rabies antibody-secreting hybrid cells reacted with all 3 rabies strains ERA, CVA and HEP-Flury in the virus neutralization, cytotoxicity and membrane immunofluorescence test; 15 reacted with the ERA and HEP strains, 2 with ERA and CVS, 3 only with ERA and 1 only with the HEP virus. 28 hybrid cultures produced antibodies that reacted with nucleocapsids from all 3 virus strains; and of these 28, only 22 reacted with nucleocapsids and no other virus components.

An analysis of antigenic relationships between street and fixed rabies virus strains was carried out on 10 additional strains of different geographic origins. The hybridomas were made with ERA as described above. The results shown in Table 8 show that the fixed Kelev virus strain was neutralized only by the antibodies excreted by Hybridoma (No. 120), and the recently discovered South African street virus (Duvenhage) only by the antibodies secreted by 2 Hybridomas. In contrast, all street virus strains, except the AF strain, which did not contain the antibody from No. 193 appeared to respond, 55 cross-reactions in the UN test. It was found that hybridoma antibodies were largely heterogeneous in the VN test with fixed virus strains. For example, the two PM and CVS strains formed from the pasteur strain reacted differently to the pasteur strain than 60 to each other. Conversely, the SAD virus and the ERA derived from it reacted in the same way with antibodies secreted by the same hybridomas. Finally it was found that the non-virulent HEP with the Kelev strain at VN and with No. 120 antibodies reacted crosswise. In addition, HEP was neutralized by antibodies that were secreted by 3 other hybridomas.

65

7

640 735

Table 8

Cross-reaction between rabies virus strains of different origins with hybridoma antibodies in the neutralization test

Origin of the strains neutralization index (log)

Land Prototype Ab- 101 110 194 120 103 193 104

comer

Fixed

Wild

SAD

> 3.0

> 3.0

> 3.0

2.5

3.0

1.5

0

dog

United States

ERA

> 3.0

3.0

> 3.0

2.5

3.0

1.0

0

Pasteur

> 3.0

> 3.0

> 3.0

1.0

3.0

1.0

0

cow

France

PM

0

> 3.0

> 3.0

3.0

3.0

0

0

CVS

> 3.0

2.0

> 3.0

0

0

0

0

human

United States

HEP Flury

> 3.0

> 3.0

0

2.0

3.0

0

0

dog

Israel

Kelev

0

0

0

> 3.0

0

0

0

dog

United States

NYC

> 3.0

3.0

3.0

2.0

> 3.0

1.0

0

bat

United States

UD

1.5

> 3.0

> 3.0

2.0

> 3.0

1.5

0

Brazil

DR

> 2.0

1.5

> 2.0

1.5

1.0

1.0

0

Fox

France

AF

2.0

2.0

> 2.0

2.0

2.0

0

0

dog

Rwanda

RD

> 3.0

> 3.0

> 3.0

2.5

> 3.0

2.5

0

human

South Africa

Duvenhage

0

0

1.5

1.5

0

0

0

Intracytoplasmic fluorescence after staining in the presence of hybridoma 104 medium

Cells infected with all viruses, regardless of their origin, showed intracytoplasmic fluorescence after fixation and exposure to Hybridoma-104 (NC test) antibodies, although none of the viruses reacted with the same hybridoma in VN. This confirms that although the hybridoma antibodies can easily distinguish fixed rabies virus strains in VN, CT and MF tests, the antibodies react with all rabies strains in the NC test.

The results shown in Tables 7 and 8 show that the anti-rabies virus antibody-producing hybrid cells are of great use for research and analytical purposes.

Hybrid cultures that secrete rabies virus neutralizing antibodies protect mice against the lethal effects of rabies virus. Cells from hybrid mass cultures C-1 and B-1 and from clone 110-5 (see Table 9) were implanted subcutaneously in BALB / c mice. The serum drawn from these mice within 2 weeks after implantation of the C-1 and 110-5 hybridomas showed a rabies antibody concentration 100 times higher than in the medium from tissue cultures of the same hybridomas. Five days after the hybridomas were implanted, mice were given a lethal dose of PM rabies virus intracerebrally. Table 9 shows that mice implanted with VN antibody-secreting hybridoma cultures survived the viral injection, whereas mice that were not implanted with VN antibody-secreting hybridomas were not protected.

Table 9

Protection of mice against rabies virus infection by hybridoma antibodies

Hybridoma number of cells VN ratio of vaccinated protected to infected mice

C-l

5 x 10 «

+

6/6

110-5

5 x 105

+

9/10

5 x 10 *

+

6/6

B-l

5 x 10 «

-

0/6

-

0

-

0/6

In this invention, the cell line consists of a hybrid culture that has the characteristics of both normal spleen and parental myeloma cells, and appears to have arisen from a single fusion event. The cells are naturally hybrid because:

30 (a) the cell line was grown for several months in selective HAT medium that inhibits the growth of parental P3 x 63 Ag8 myeloma cells, but not that of normal spleen cells, which in turn are likely to be no more than 4-5 weeks would survive in vitro; 35 (b) The number of chromosomes in the hybrid cells is almost equal to the sum of the parent chromosomes of the normal spleen and myeloma cells;

(c) The HK-PEG-1 hybrid not only produces IgGl, which is also excreted by P3 x 63-Ag8, but

40 also IgG3, and probably hybrid molecules; and

(d) The hybrid produces antibodies with antivirus activity, but P3 x 63-Ag8 does not.

The antibodies are best grown by multiplying the hybrid cell lines in vitro. However, antibodies can also be produced after injection of the hybrid cell lines in tissue-compatible animals, so that the antibodies are formed in vivo. So z. B. mice, relatively inexpensive animals, are injected with the mouse-derived antibody-producing cells of the present invention to obtain isolatable antibodies in the sera and ascitic fluids.

Table 6, described above, shows the amount of antibody available under these circumstances.

55 The antibodies produced by the hybrids can be obtained by conventional methods and used in analytical medical research to identify virus antigens in blood samples and culture media. These in vitro generated antibodies are homogeneous and specific to the virus antigen. A stock of different homogeneous antibodies, each very specific to a particular antigen, allows the scientist to immediately determine the specificity of an antigen and / or to characterize virus antigens. Antibodies can also be given to sick animals to help fight the disease.

s

Claims (26)

640 735 2nd PATENT CLAIMS 1. A method for producing an antibody-producing continuous cell line, consisting of the preparation of a cell hybrid fused from a virus antibody-producing and a myeloma cell and cultivation of this hybrid. 2. The method of claim 1, wherein the hybrid is grown in vitro. 3. The method of claim 2, wherein the hybrid is cloned and grown in a medium containing hypoxanthine-aminopterin-thymidine. 4. The method of claim 1, wherein the hybrid is injected into a histocompatible animal and bred in vivo. 5. The method of claim 4, wherein the hybrid consists of a clone. 6. The method of claim 1, wherein the virus antibody-producing cell is a cell selected from spleen cells and lymph node cells. 7. The method of claim 5, wherein the virus antibody-producing cell is a spleen cell. 8. The method of claim 6, wherein both the spleen and the myeloma are from a mouse. 9. The method of claim 7, wherein the mouse is a BALB / c mouse. 10. The method of claim 9, wherein the myeloma (P3 x 63 Ag8) is from the MOPC-21 line. 11. The method of claim 1, wherein the virus antibody-producing cell is taken from an animal immunized with virus. 12. The method of claim 2, wherein the virus antibody-producing cell is taken from an animal immunized with virus. 13. The method according to claim 7, wherein the virus antibody-producing cell is removed from a mouse immunized with virus. 14. The method of claim 12, wherein the virus is an influenza virus. 15. The method of claim 12, wherein the virus is a rabies virus. 16. The method according to any one of the preceding claims, wherein the continuous cell line consists essentially of the HK-PEG-1 clone. 17. A method for producing an antibody-producing continuous cell line according to claim 1, consisting of injecting a BALB / c mouse with virus to produce antibodies by mouse spleen cells, forming a fused cell hybrid of the spleen cells producing virus antibodies with one of the MOPC line -21-derived BALB / c myeloma cell clone (P3 x 63 Ab8), and cultivation of the hybrid in vitro in selective HAT medium. 18. The method of claim 17, wherein the virus is an influenza virus. 19. The method of claim 17, wherein the virus is the 6-94 influenza virus strain. 20. The method of claim 17, wherein the virus is simian virus 40. 21. The method of claim 17, wherein the virus is a rabies virus. 22. Antibody-producing continuous cell line, produced according to the method of claim 1. 23. Continuous cell line according to claim 22, which produces influenza antibodies in vitro in hypoxanthine-aminopterin-thymidine medium and consists of a fused cell hybrid of spleen cells of a BALB / c mouse pre-infected with an influenza virus and MOPC-21 mouse myeloma cells. 24. Continuous cell line according to one of claims 22 and 23, consisting essentially of the HK-PEG-1 clone. 25. A method for producing virus antibodies, consisting of culturing a continuous cell line produced according to the method of claim 1 in a culture medium and collecting the virus antibodies. 26. A method for the production of antibodies according to claim 25, consisting of culturing one according to the lo drive according to claim 17 produced continuous cell line and collection of the antibodies produced. 15