JPH0320238B2 - - Google Patents

Description

[Detailed description of the invention]

(a) Industrial application field The present invention is directed to the varicella zoster virus (Variceila
This article relates to a human monoclonal antibody (hereinafter referred to as MCA) that recognizes the 90K and 55K glycoprotein antigens of Zoster Virus (hereinafter referred to as VZV) and its production method. The purpose is to provide a human-type MCA that is specific to a glycoprotein antigen of a specific molecular weight of VZV, which is useful in the diagnosis and treatment of viral infections caused by VZV. (b) Conventional technology and its problems A mouse-type MCA for VZV has already been produced (T. Okuno et al.
Varology, vol. 129, p. 357, 1983). However, since mouse-type MCA is antigenic to humans, antibodies are generated when it is administered into the human body, causing anaphylaxis-like side effects. Therefore, humanized MCA is particularly desirable for therapeutic purposes. Human-type MCA includes MCA for cancer cells (J. Schlom et al., Proc.
Natl.Acad.Sci.USA, Volume 77, Page 6841,
1980), MCA against technus toxin (N, N.
Teng et al., Proc. Natl, Acad, Sci. USA, vol. 80,
7308 pages, 1983), MCA for eczema (CA
Croce et al., Nature, vol. 288, p. 488, 1980). However, mouse-type MCA
Compared to VZV, only a limited number of human MCAs have been known so far, and the human MCA against VZV is also unknown. There are two reasons why human-type MCA is more difficult to produce than mouse-type MCA. The first point is that the efficiency of forming MCA-producing hybridomas is very low when human lymphocytes are used, and the second point is that it is difficult to immunize humans in many cases, so it is difficult to effectively immunize humans. It is difficult to obtain stimulated human lymphocytes. The latter issue is particularly important in producing human MCAs specific for infectious diseases or cancer. (c) Means for solving the problem The present inventors believe that, since it is currently impossible to administer VZV to humans to immunize them, a method of sensitizing human lymphocytes outside the body is essential. As a result of much thought and research, we decided to create a hybridoma by fusing human lymphocytes (antibody-producing cells) sensitized with VZV with tumor cells such as mouse myeloma cells or human lymphocyte cell lines. Alternatively, by transformation with Epstein-Barr Virus (hereinafter referred to as BEV),
We obtained a cell line that produces VZV-specific human MCA. The present inventors then cultured such cell lines and were able to obtain human MCAs that specifically reacted with the 90K and 55K glycoprotein antigens of VZV from the culture. That is, the present invention is a human MCA that specifically reacts with the 90K and 55K glycoprotein antigens of VZV. Such humanized MCA is produced by fusing tumor cells with human lymphocytes sensitized with VZV or a protein or glycoprotein derived from the virus in the presence or absence of mitogen in vitro. It can be obtained by creating a hybridoma that produces human-type MCA that reacts with , culturing the hybridoma and/or a cell line derived therefrom, and collecting MCA from the culture. Alternatively, humans sensitized as described above may be
Also from cultures of lymphoblastoid cells that produce human-type MCA that specifically reacts with VZV, obtained by transforming lymphocytes with EBV, and/or cell lines derived therefrom. can get. In the present invention, human lymphocytes include, in addition to antibody-producing B lymphocytes, their precursor cells, memory cells, and the like. These cells are obtained from tissues such as human spleen, lymph nodes, peripheral blood, bone marrow, tonsils, and adenoids. However, there are extremely few B lymphocytes specific for a given antigen, and even if hybridomas are directly produced or transformed with EBV, the probability of obtaining cells that produce MCA specific to that antigen is extremely small. Therefore, in the present invention, the following lymphocytes are in vitro treated with antigens.
sensitize. In in vitro sensitization, for example, lymphocytes are dispersed in a culture medium at a cell density of 0.5 x 10 ⁶ to 3 x 10 ⁶ /ml, and 0.1 to 0.2 ml each is placed in a 96-well plate, or 0.5 to 2 ml is placed in a 24-well plate. Put them in one by one. As an antigen, the infectivity has been inactivated.
Using VZV or its virus-derived protein or glycoprotein, 0.2 ng/ml to 1000 ng/ml, preferably 1 to 100 ng/ml of antigen is added to the lymphocyte suspension. Lymphocyte mitogen is also effective for antigen sensitization. Porkweed mitogen (hereinafter referred to as mitogen)
PWM), protein A, and B cell growth factor (so-called BCGF). When using PWM, 1 μg/ml to 200 μg/ml, preferably about 20 μg/ml, is added to the lymphocyte suspension. In particular, when an antigen and PWM are added together, the number of lymphocytes specific for that antigen synergistically increases. In this way, antigen or antigen and mitogen are added to the lymphocyte suspension, and the mixture is incubated at 37°C for 3 to 20 days, for example.
Preferably, the culture is carried out for 4 to 8 days. Various serums can be used as the culture medium, such as fetal bovine serum (hereinafter referred to as
FCS), a culture medium containing calf serum, horse serum, etc. (e.g. RPMI1640 or Zarbetsko MEM)
is used. The sensitized human lymphocytes thus obtained are fused with mouse or human lymphoid cell lines or
Transformed by EBV. Cell fusion partners include mouse myeloma cell lines,
P3×63Ag8, P3−NS1/1−Ag4−1, P3×
63Ag8u1, SP2/0Ag14, P3×63Ag8.6.5.3,
MPC1145.6, TG1.7 or SP-1 is used. As a human lymphoid cell line, myeloma
RPMI8226 strain, lymphoblastoid cell u729-6 strain,
ARH77 strain and also B lymphoma are also used.
In addition, heteromyeloma (NNTeng) derived from mouse myeloma and human lymphoid cells
80, page 7308, 1983) can also be used. The sensitized lymphocytes and the mouse or human lymphoid cell line are fused according to a known method as follows. For example, the ratio of sensitized lymphocytes and lymphoid cell lines is 10:1 to 1:100, preferably 1:1.
Mix at a ratio of 1 to 1:20, add a suitable cell fusion solution, such as RPMI1640 containing 35% polyethylene glycol (molecular weight approximately 1000-6000) and 7.5% dimethyl sulfoxide, and incubate at room temperature to 37°C. Shake for several minutes, then gradually dilute with RPMI1640 and then RPMI1640 supplemented with 10% FCS. After washing, use hypoxanthine-aminopterin-thymidine (hereinafter referred to as HAT) selection culture medium to reach a cell density of 0.5 to 5 x ^105. Prepare the amount of cells/ml. Dispense 0.2 ml of this into a 93-well plate, and add 5% CO2 _.
Culture at 35-38° C. for 2-4 weeks in humid air containing water. In the HAT culture solution, only hybridomas survive, and unfused 8-azaguanine-resistant lymphoid cell lines and lymphocytes die. After culturing, the antibody titer in the culture solution is screened to select hybridomas producing the desired MCA. Screening methods include radioimmunoassay (RIA), enzyme-linked immunosorbent assay (ELISA),
There are fluorescent antibody methods, etc. Instead of producing hybridomas, sensitized lymphocytes are transformed with EBV and a cell line that permanently produces MCA can be obtained.If cells that continue to produce EBV, such as the B95-8 cell line, are cultured, The culture supernatant contains infectious EBV. This culture supernatant is used as a source of EBV to transform lymphocytes. When transforming the EBV, e.g. 0.01
Add ~10, preferably 0.2-1 transform units ( _TD50 )/lymphocyte. For example, after incubating at 37°C for about 1 to 2 hours, free virus is removed by centrifugation, and the resulting EBV-infected cells are added with 10 to 20% FCS.
Disperse in RPMI1640 medium. Seed 0.2 ml of this cell suspension (cell density: 400 to 50,000 cells/ml) in a 96-well plate. After culturing for 3 to 5 weeks at 37° C. in a CO ₂ incubator, the supernatant is screened in the same manner as for hybridomas. The MCA-producing cell line thus obtained is cloned by the limiting dilution method or the soft agar method. The cloned hybridoma or EBV transformant is stored in a cell storage tank containing liquid nitrogen. When such a hybridoma or EBV transformant cell line (cell line) and/or a cell line derived therefrom is cultured in large quantities by an appropriate method, the human type targeted by the present invention can be obtained from the culture supernatant.
You can get an MCA. It is also possible to transplant this hybridoma into an animal to form a tumor, and obtain human-type MCA from the ascites or serum. Characteristic of the present invention obtained by such a method
MCA specifically reacts with VZV. In particular, human MCAV1 has a molecular weight of 108K, 105K, and 62K.
Human MCAV2 recognizes glycoprotein antigens with molecular weights of 90K and 55K. Then, in the presence of complement for both V1 and V2, VZV was injected at a concentration of MCA antibody of 1 μg/ml or less.
It has excellent sanitizing activity that can inactivate 50%. Hereinafter, the present invention will be explained in detail with reference to Examples. (d) Examples Hybridoma producing human MCA for VZV (1) Preparation of VZV antigen VZV in culture medium of human fetal lung cells (HeL)
(Kawagucni strain) was inoculated, and when cell degeneration appeared, the cells were removed with a rubber policeman. Cells were disrupted by sonication for 1 minute and then centrifuged at 3000 rpm for 10 minutes. The supernatant was placed on a layer of cesium chloride and centrifuged at 23,000 rpm for 3 hours. VSV clustered between densities 1.4 and 1.2. This virus fraction was further heated at 25,000 rpm.
The pellet was precipitated by centrifugation for 1.5 hours, and the precipitate was dispersed in phosphate buffered saline and sonicated three times for 5 seconds. The virus preparation thus obtained was inactivated by ultraviolet irradiation and then used in the following experiments. (2) Sensitization of lymphocytes by VZV Human tonsil lymphocytes were cultured in medium A (RPMI1640+
20% fetal bovine serum + 20mM HEPES + 2mM glutamine + 1mM Napyruvate + 0.02mg/
ml serine + 80 μg/ml gentamicin) to 2.6×
10 ⁶ cells/ml were suspended. Add to this the VZV obtained in (1) above (final concentration 17 ng protein/ml) and
PWM (20 μg/ml) was added and cultured for 3, 5, or 7 days. The thus sensitized lymphocytes were used for the following cell fusion. (3) Cell fusion with mouse myeloma cell line P3×63Ag8U1 (abbreviated as P3U1).
The cells were cultured in calf serum + 2mM glutamine + 80μg/ml gentamicin). The cell concentration at the time of use was 6×10 ⁵ cells/ml. 4 groups of sensitized lymphocytes (3 wells together) from (2) above and P3U1
were each washed twice with serum-free RPMI1640. Lymphocytes from each group and 5×10 ⁶ P3UI were combined in a test tube. Centrifugation was performed at 1500 rpm for 5 minutes, and the supernatant was discarded. The cell pellet was well dispersed by tapping the test tube. Add 0.5ml of polyethylene glycol solution (RPMI1640 5.75ml + polyethylene glycol)
1000 3.5ml + dimethyl sulfoxide 0.75ml)
(abbreviated as PEG solution) was added to gently suspend the cells. After 1 minute, add 0.5ml RPMI1640, after another minute, add 1ml RPMI, and after another 2 minutes, add 4ml HAT culture solution (RPMI1640 + 20% fetal bovine serum + 80μg/ml gentamacin +
95 μM hypoxanthine + 0.4 μM aminopterin + 1.6 μM thymidine), and after another 2 minutes, 4 ml
HAT culture solution was added. Finally, a 25 ml cell suspension was made with HAT culture medium. This was plated on one culture plate (96 wells) and cultured at 37° C. in air containing 5% CO ₂ . Every week, add half of the culture solution to new HT culture solution (HAT with A removed).
I exchanged it and got a hybridoma. (4) Measurement of human anti-VZV antibodies The culture supernatant of the hybridoma thus formed was tested with 1gM or 1gG anti-VZV antibody.
MCA was measured by enzyme antibody method. That is,
VZV (Kawaguchi strain) to measure anti-VZV antibodies
2 μg protein/ml was each immobilized on a 96-well plate of Falcon Microtest. 60μ of hybridoma culture supernatant was added to this plate and left at room temperature for 1 hour. and 1%
Wash with Hank's salt solution (HBSS-BSA) containing BSA (bovine serum albumin), then add 60μ of goat anti-human 1gG antibody or anti-1gM antibody-alkaline phosphatase (2000-fold dilution) and Allowed time to react. Further HBSS
- After washing three times with BSA, 100 μ of a solution of P-nitrophenyl phosphate dissolved in a 1 M diethanolamine + 1 mM MgCl ₂ solution at pH 9.8 at a ratio of 0.6 mg/ml was added. From 30 minutes
After 60 minutes, the absorbance at 405 mμ was measured. The results are shown in Table 1.

[Table] Example 2 Lymph was isolated from a spleen surgically removed from a patient with autoimmune thrombocytopenia by the Ficoll-Pack method. The lymphocytes were treated with VZV (17ng/ml) and PWM in the same manner as in Example 1 (2).
(20 μg/ml) and sensitized for 6 days. The thus sensitized lymphocytes were subjected to cell fusion in the same manner as in Example 1 (3). Regarding the hybridoma culture supernatant thus formed, total human 1gG and 1gM, 1gG and 1gM reacting with VZV-infected Hel cells,
1gM, plus 1gG and
1 gM was measured by enzyme antibody method. The results are shown in Table 2 as the number of corresponding holes in a 96-well plate. Table 2 shows that production of 1gG type MCA selective to VZV occurs. Furthermore, it can be seen that the number of hybridomas producing such a VZV-selective MCA significantly increases upon sensitization with VZV+PWM. Although such hybridomas appear even with PWM alone (6/55),
There is an increase in non-selective hybridomas (9/27) compared to VZV-only sensitization.

[Table] Example 3 Cloning of anti-VZV antibody-producing hybridoma and specificity of anti-VZVMCA. (1) Cloning Take out cells from the anti-VZV activity positive hole,
Count the number of cells, 1 cell/well or 10 cells/well.
Cells were seeded in a 96-well culture plate. After 3 to 4 weeks, the cells had proliferated sufficiently, so the culture supernatant was screened for anti-VZV antibody activity. In this way, the derived cells were selected from the one that was seeded as much as 1 cell/well. This operation was repeated 2 to 4 times to ensure that all clones had anti-VZV antibody activity. Three hybridomas were established through mouse x human cell fusion. Those hybridomas are 6
MCA was produced at 25 μg/day/10 ⁶ cells. These were named V1, V2, and V6. (2) Specificity of MCA The subclasses and L chain types of V1, V2, and V6 were determined. Sheep anti-human 1gG, anti-human 1gG2,
The subclass was determined by the Ochterloni method using anti-human 1gG3 and anti-human 1gG4. As a result, V1, V2, and V6 reacted only with the anti-human 1gG1 antibody. Furthermore, ELISA using alkaline phosphatase goat 1gG anti-human K chain or anti-infection chain
As a result, it was found that all three MCAs had strands. To examine the immunospecificity of V1, V2, and V6,
ELISA was performed using various viruses or their host cells as antigens. The results are shown in Table 3. V1, V2, and V6 reacted to all six strains of VZV tested, but did not react at all to the host cell HEL. Also, V1 is HSV1 type and 2
It reacted weakly to the type, but V2 and V6 did not react. Furthermore, neither MCA responded to CMV or EBV.

【table】

[Table] Example 4 Determination of antigen molecules recognized by anti-VZV/MCA Antigen molecules were determined by immunoprecipitation and nylium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-
PAGE). A 1/5 volume of human infant lung cells (HEL) suspended with trypsin was added to a monolayer culture (64 cm ² ) of human infant lung cells (HEL) on the second day of culture.
VZV-infected cells were inoculated. When 80% or more cell degeneration (CPE) appeared after 48 hours, the ground was injected with 4 ml of 100 μci tritium glucosamine hydrochloride ([ ^1.6-3 H(N)] glucosamine; 20 ci/mmol;
1/10 Glucose MEM containing (NEN Research Products, Boston, MA) - 2% FCS or 5 ml of
50μci〔 ^35S 〕-Methionine (400ci/mmol; NEN
Research Products) containing 1/10 methionine
It was replaced with MEM-2%FCS. 18 further at 37℃
Cells were cultured for an hour, during which time cells were collected when CPE had spread throughout the cell monolayer. Wash the collected cells,
10mM Tris HCl (PH7.4), 0.15MNaCl, 1mM
Phenylmethylsulfonyl fluoride (Signa
Chemicals CO., ST Louse, Mo), 1mM ethylenediaunetetraacetic acid, 1% Tritton X-100,
It was dispersed and solubilized in 0.1% sodium dodecyl sulfate and 1% sodium deoxycholic acid (RIPA buffer) by sonication. This was layered on a 60% sucrose solution, and insoluble matter was removed by ultracentrifugation at 100,000g for 1 hour, and the supernatant was used as an antigen at -70°C.
It was stored frozen. [ ^3H ]-glucosamine-labeled antigen fraction (200μ,
Mix 400,000 cpm) or [ ^35S ] methionine-labeled antigen fraction (200μ1200000cpm) with 150μ of hybridoma supernatant, and incubate at room temperature for 30 minutes and then at 4℃.
The mixture was allowed to react overnight. The next morning, add previously swollen Protein A-Sepharose CL4B beads (3 mgr in 100μ suspension: Pharmacia
Fine Chemicals, Inc., Tokyo) and reacted at 4°C for 3 hours. Beads containing immune complexes were pelleted by centrifugation and washed five times with 2 ml of RIPA buffer and twice with 2 ml of 10 mM Tri HCl (PH 6.8). After washing, the immune complexes were washed with 110μ
SDS-sample buffer was added and eluted at 100°C. Electrophoresis of SDS-PAGE solubilized samples was performed at room temperature.
It was carried out according to the standard method of Tris HCl-glycine-SDS buffer system (PH8.3). slab gel
After immersion in 1M sodium salicylate aqueous solution,
Dry on paper. The dried gel was exposed to KODAKX-OMATS film at -70°C for 7 to 21 days. The results are shown in Figure 1. V1 of MCA is molecular weight
It was recognized that V2 recognized glycoproteins with molecular weights of 108K, 105K, and 62K, and different glycoproteins with molecular weights of 90K and 55K. In addition, in Figure 1, V1 (column A) and V2 (column B
column) was precipitated with [ ^35S ]-methionine-labeled viral antigen. V1 (column C) and V2 (column D) are [ ³ H]-
Precipitated with glucosamine-labeled antigen. The arrow on the right indicates the mobility of the standard protein. Also, K
means kilodalton. Furthermore, V6 showed exactly the same electrophoretic pattern as V1. Example 5 Virus neutralizing activity of anti-VZV/MCA Culture supernatants of anti-VZV/MCA producing cells were collected,
MCA was precipitated with 50% saturated ammonium sulfate and dialyzed against phosphate-buffered saline. MCA this
It was used as a standard sample in the following experiments. The amount of MCA contained in these preparations was determined by a one-dimensional immune diffusion method using an anti-human 1gG antibody. The MCA preparation was serially diluted 3 times with 5% FCS-MEM, and the solution (200μl) and VZV (Kawaguchi strain, 4000PFU, 100μ) were added.
and guinea pig complement (5-fold dilution, 100μ) or 5% FCS-MEM (100μ),
The cells were cultured at 37°C for 1 hour. This virus mixture is
The virus was added to the previously cultured monolayer culture of HEL cells and allowed to adsorb at 37°C for 1 hour. Then 5% FCS- containing 2.5% methyl cellulose
MEM was overlaid and cultured in a CO ₂ incubator for 5 to 7 days. Thereafter, infected HEL cells were fixed with 10% formalin, further stained with 0.15% crystal violet, and the number of plaques was counted. As a result, the virus activities for MCA, V1, V2 and V6 were as shown in Table 4. It can be seen that V1 and V2 have excellent neutralizing activity in the presence of complement, with the amount of MCA required to inactivate 50% of the virus being less than 1 μg/ml.

[Table] Example 6 Preparation of human x human hybridoma Human spleen lymphocytes were cultured in culture medium A ((2) of Example 1).
(see) at a density of 1.2 x 10 ⁶ cells/ml. Each 1.2 ml of this cell suspension was placed in a well of a 24-well plate. There, the antigen (VZV, Kawaguchi strain)
Add to 4.9ng protein/ml, and
PWM was added at a concentration of 20 μg/ml. _CO2
The lymphocytes were sensitized to the antigen by culturing in an incubator for 6 days. Human lymphoblastoid cells 729-6 were previously cultured in culture medium B (RPMI 1640 + 10% FCS + 2mM glutamine + 80 μg/ml gentamicin). The cell density at the time of use was 8×10 ⁵ cells/ml.
The above sensitized lymphocytes were pooled for 12 wells and serum-free.
Washed twice with RPMI1640. After washing the 729-6 cells twice, 2×10 ⁷ 729-6 cells were mixed with the sensitized lymphocytes in a centrifuge tube. Centrifugation was performed at 1500 rpm for 5 minutes, and the supernatant was discarded. The cell pellet was well dispersed by tapping the tube. 2ml of this
PEG solution was added to gently suspend the cells. After 1 min, add ml RPMI1640, after another 1 min 4 ml RPMI, after 2 min 8 ml HAT culture (RPMI 1640 + 20% FCS + 80 μg/ml gentamicin + 95 μM hypoxanthine + 0.4 μM aminopterin + 1.6 μM thymidine), and after another 2 min 16
ml of HAT culture solution was added. Finally, 100 ml of cell suspension was prepared using HAT culture medium. This was plated on four culture plates (96 wells) and cultured at 37° C. in air containing 5% CO ₂ . Hybridomas were obtained by replacing half of the culture medium with fresh HT culture medium every week. After 3 to 4 weeks of culture, antibodies were measured. Hybridoma growth was observed in 118 out of 384 wells on 4 plates. Antibiotics of these culture supernatants
As a result of screening for VZV antibody activity, 16 wells were positive. Antibody titers against uninfected HEL were not measured in these wells, and the human MCA produced is considered to be VZV-specific. All isotypes were 1gG. Example 7 Preparation of transformed cells using EBV (1) Preparation of EBV A persistent EBV producing strain derived from human B cells.
Return the B95-8 cell line from freezing and use culture medium C.
(RPMI1640 + 10% FCS + 50 I.U./ml streptomycin + 50 μg/ml penicillin G potassium) at a final concentration of 2 x 10 ⁵ cells/ml at 37°C.
The cells were cultured for 3 days. After that, the final concentration is 2× again.
The cells were inoculated into new culture solution C at 10 ⁵ cells/ml and cultured for an additional 2 days. B95-8 cells in the logarithmic growth phase scaled up in this way were added to a final concentration of 1
×10 ⁵ cells/ml and further at 37°C in fresh culture medium C.
Culture for 7 days and remove the culture supernatant containing EBV.
Centrifugation was performed at 3000 rpm for 30 minutes, and the mixture was further filtered through a Millipore filter with a pore size of 0.45 μm. The EBV preparation solution thus obtained was stored frozen at -80°C. The titer of EBV in the preparation solution was 1×10 ⁵ TD ₅₀ /ml. (2) Lymphocyte sensitization by VZV and lymphocyte transformation by EBV Human spleen lymphocytes were cultured in culture medium B (Example 1).
(3)) were suspended at 2×10 ⁶ cells/ml. 1.2 ml of this cell suspension was placed in each hole of a 24-well plate. This includes VZV (Kawaguchi strain) 4.9ng protein/ml
was added and cultured in a CO ₂ incubator for 4 days. The thus sensitized lymphocytes were centrifuged at 15,000 rpm for 5 minutes, and the cell pellet was well dispersed by tapping the test tube. In addition to the above
The EBV preparation obtained in (1) was added at a volume of 0.5 TD ₅₀ /cell and incubated at 37° C. for 60 minutes with occasional stirring. Finally, the culture solution
DRPMI1640+10%FCS+2mM glutamine+
Add 1mM Na-pyruvic acid + 0.02mg/ml serine + 80μg/ml gentamicin), plate the cells in a 96-well plate at 1.2 x ¹⁰⁴ cells/well/0.2ml culture medium, and culture at 37°C with 5% _CO2. Cultivated in air. Half of the culture solution was replaced with new culture solution D every week to obtain lymphoblastoid cells. Lymphoblastoid cells grew into the wells of all 4 plates. 4 weeks after transformation
As a result of measuring the VZV antibody titer, 238 out of 384 wells had antibodies. However, before cloning began, many cells stopped producing antibodies and continued to produce antibodies. Twelve clones were cloned by soft agar method.
Three of them were established as lymphoblastoid cell lines producing anti-VZV human-type MCA.

[Brief explanation of the drawing]

Figure 1 shows the antigens recognized by anti-VZV and MCA.
It is an SDS-PAGE electropherogram.

Claims (1)

[Scope of Claims] 1. A human monoclonal antibody that recognizes glycoprotein antigens of the varicella-zoster virus with molecular weights of 90K and 55K. 2. In vitro, human lymphocytes sensitized with varicella zoster virus or proteins or glycoproteins derived from the virus are fused with tumor cells in the presence or absence of mitogenes to produce varicella zoster. It consists of creating a hybridoma that produces a human monoclonal antibody that recognizes glycoprotein antigens with molecular weights of 90K and 55K of the virus, culturing the hybridoma and/or a cell line derived from it, and collecting the monoclonal antibody from the culture. , a method for producing human monoclonal antibodies that recognize glycoprotein antigens of the varicella-zoster virus with molecular weights of 90K and 55K. 3. A method for producing a human monoclonal antibody that recognizes glycoprotein antigens of the varicella-zoster virus with molecular weights of 90K and 55K, as set forth in claim 2, wherein the tumor cells are mouse myeloma cells. 4. A method for producing a human monoclonal antibody that recognizes glycoprotein antigens of the varicella-zoster virus with molecular weights of 90K and 55K according to claim 2, wherein the tumor cells are human-derived lymphoid cell lines.