JPH06506947A - Antivirus hybrid antibody - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] Background of the invention The present invention provides recombinant hybrid components for use in the treatment and prevention of viral infections. Concerning children.

There is a wide range of foreign substances and organisms that can enter the body and cause disease. child In response to such intrusion, mammals, including humans, respond with an "immune response." , this is the result of numerous complex interactions that occur between various cells and humoral factors. It is. Many different cells are involved, but only one is involved in generating an immune response. The lymphoid cells are the main cells, and they protect against foreign substances such as bacteria, viruses, and foreign cells. protect the body.

There are two main classes of lymphocytes, called B cells and T cells. each club The cells are formed from original hematopoietic stem cells.

Mature T cells are divided into three subclasses based on the different actions they perform. It is.

Helper T cells (Th) have the function of promoting or enhancing B cell antibody production.

They are called cytotoxic killer T cells (Tk) or cytotoxic class 1926 (CTL). Those that are used directly kill cells by lysing them. Suppressor T Different subclasses of T cells (Ts) suppress or reduce the immune response. ras expresses a variety of cell surface proteins, some of which are characterized by specific subclasses. There is something called a "marker protein". For example, most Th cells CD4 protein is expressed on the cell surface, whereas most CTL and Ts cells express the CD4 protein on the cell surface. Expresses C″D8 protein on the surface. Swain r different action analysis activities The fact that there are two distinct classes of mouse B-cell growth factors depending on gender (E vidence for two distinct classes of Murine B Ce1l Growth Factors with Ac tfvities in Dffferent Functfonal As5 ays) J, Exp, Med, 158:822 (1983) o Furthermore, Mature T cells have T cell receptors (TCR) on the cell surface, so they are different from immature cells. This TCR is found on mature T cells, although it can be differentiated Involvement of self-antigens, which are membrane-permeable protein complexes and encoded by MHC genes. It has the function of recognizing antigens.

As we currently know, the initiation and maintenance of an immune response relies on cell-to-cell interactions. present on the surface of B cells, T cells, foreign substances, foreign cells, and infected cells. , which relies on the recognition of specific proteins or protein complexes and the interactions between them. Ru.

There are at least two distinct types of immune responses: cell-mediated responses and antibody-mediated responses. There are aspects of Both begin when a T cell recognizes a foreign antigen. Thin The cell-mediated response involves the cytolytic activity of activated CTLs upon encountering the antigen; Proceeds in the absence of cysts. In addition, CTLs are activated nonspecifically, and Lysing all nearby cells by binding antibodies to cell surface proteins Can be done. For an antibody-mediated response to occur, an antibody must be activated upon encountering an antigen. Th cells interact with B cells and produce known immunoglobulins or antigens. Stimulates B cell production of body fluid proteins.

T cells directly participate in cell-mediated immune responses to foreign antigens, but antibody B Cell production is the most important aspect of immunity. Various essential antibodies are produced by immunoglobulin Formed by the diversity of Brin genes. Gene rearrangement further increases the number of types To increase. Each set of mature immunoglobulin genes undergoes further gene rearrangement. This is due to this. To provide even more variety, we offer a number of products with different characteristics. There is a class of immunoglobulins. In the structure of immunoglobulins and proteins For more information, see "Gene I11" by Levin, John Wiley and Sa. (Johnν+Iey and S.

n5) See N, Y. (1987).

Advances in genetic engineering can aid the natural immune system and provide reagents for diagnostic tests. It is used for the development of For example, it is suitable for vaccines and against various antigenic determinants. The corresponding protein sequence has been prepared synthetically or by recombinant DNA technology. .

Antibodies are of great importance for diagnostic and therapeutic applications due to their diversity and specificity. . Molecular biology techniques are being used to expand the potential for scientific applications of antibodies. came. For example, B cells that produce a single type of antibody can be established and expanded to An in vitro antibody with a single specificity known as mAb J You can get the body source.

Such established cell lines are called "hybridomas."

Until recently, the origin of most mAbs was murine hybridomas. mouse mA b is widely used in diagnostic methods, but it is not accepted by mammals, including humans and non-homologous mice. Not suitable for dynamic immunotherapy or therapeutic use. Furthermore, mouse antibodies are foreign to other mammals. It is recognized as such and generates an immune response that can itself cause disease. that Therefore, human mAbs would be extremely useful in a wide range of human diseases. However, humans Production of mAbs has proven to be much more difficult than production of mouse mAbs. . Therefore, human mAbs have not yet been obtained in sufficient quantities and types to be used for therapy. .

To solve the problem of immune response to foreign mAbs and lack of suitable human mAbs, hybrid immunoglobulins, at least in part using genetic engineering techniques. It has been done to construct molecules. This molecule is the antigen-binding site of mouse antibodies. , the remaining portion consists of foreign substances and unrecognized human antibody sequences. Jones (Jo nes) et al. “Replacement of human antibody complementarity determining region with mouse antibody complementarity determining region J (Replacing the Complementality-Dete rmining Regions in a Human Antibody with those from a Mouse) ) 321:522-525 (198B), these hybrid antibodies , which sometimes generates an immune response in human therapy, and the parent mouse antibody It often doesn't work. For information on the use and background of mouse and human mAbs, see Carlsson et al., “Monoclonal antibodies in the 1990s: the path to omnipotence” As an ingredient J (Monoclonal Antibodies 1nto th e 90's: AII Purpose Tool) Bio/Technology< Bio/Technology) 7: 587-573 (1989). and.

Summary of the invention The present invention can be used for human treatment, particularly for the treatment of viral infections and for the treatment of tumors. , a novel drug and method that combines both cell-mediated and antibody-mediated immune responses in a single drug. Regarding the law.

In the treatment of viral infections, the novel agent according to the invention can be used to treat viral infections. By concentrating cytotoxic class 1928 on cells and causing lysis of infected cells. , acts to prevent viral infection.

The present invention provides hybrid antibodies, and the hybrid antibodies of the present invention are human immunoglobulins. Corresponds to the C region (Constant portion) of Robulin G a basic region and a linkage selected to exhibit specificity for a particular target antigen. It consists of a binding site and a binding site that binds and activates human CTL. This hybrid When hybrid antibodies are used to treat viral infections, preferred hybrids are: Binds to proteins involved in viral infection, i.e., viral “cell recognition” proteins It has a target antigen binding site that neutralizes the ability of the virus to infect. . Cell recognition proteins are often expressed on the surface of virus-infected cells. ,/1 hybrid antibodies can also bind to these infected cells. No＼Iburi Nodo Anti When the body binds to both virus-infected cells as well as CTLs, activation of CTLs, followed by lysis of infected cells occurs.

Human Immunodeficiency Virus (HIV): This is the cause of acquired immunodeficiency syndrome (AIDS) The hybrid antibody according to the invention used for the treatment of infections (causing CD3) The binding region that binds and activates CTLs and all HI generated from the surface of infected cells. It is desirable to have a binding region specific to the V antigen. For example, antigen recognition binding site The position is the V region (variable region) of an antibody specific to the coat protein of HIV.

Brief description of the drawing Figure 1 shows the binding site, hinge region, L chain (short chain) and H chain (long chain), and their Diagram of an immunoglobulin molecule shown in a Y shape, consisting of V and C regions corresponding to It is.

Figure 2 is a diagram of a protein complex, and this complex is linked to viruses and virus-infected cells. , or a single immunoglobulin binding site capable of recognizing viral antigens and CD Yet another single immunoglobulin binding that can recognize and bind to 3 to activate CTLs. and their binding sites in the immunoglobulin C region (constant do The immunoglobulin hinge is separated from the CH2 and CH3 regions (sain). The immunoglobulin hinge part is separated from the region.

Figure 3 is a schematic diagram of the DNA structure containing the VH-D-J gene. A method for cloning regions within a single recombinant protein complex and cell-mediated reactions By combining this and antibody-mediated reactions, It has been found that a new effective therapy can be obtained.

INDUSTRIAL APPLICABILITY The present invention is useful for the treatment and prevention of human viral infections, and It concerns hybrid antibodies produced by this technology.

At the heart of the hybrid antibodies of the invention are at least a portion of human immunoglobulin G There is a basic domain of (IgG). As shown in Figure 1, IgG has two identical H! IHSH' and four proteins formed by two identical light chains LSL' It is a complex. These chains are connected by disulfide bonds and form a Y-shaped complex. I'm reading. However, in solution the molecule assumes a more spherical morphology.

Seafence analysis of immunoglobulin proteins has revealed specific regions within each of these chains. It has become clear that there are areas or functional areas. Each chain has an amino terminal It has V regions (VL and VH). V created by the combination of VL and VH The region constitutes the antigen recognition site or "binding site" of the molecule. 2 per molecule There are two binding sites.

The V regions of these chains are highly diverse in sequence, providing a variety of antibody binding sites and Exhibits specificity for antigens. Furthermore, each chain essentially has a C region (const ant region), and this C region is recognized by the binding site. It does not change depending on the nature of the antigen. The L chain has a single C region (con There is a stunt region (CL), but on the other hand, the H chain has three mutually different regions. C region (constant region) (CH1.

CH2, CH3). The pair of CL and CHI is the first C domain (consta nt doiain) forms CI, and the pair of CL and CH2 forms the second C domain C2 is formed, and the combination of CL and CH3 forms the third C domain C3. 4 pieces The C domains, the two C1, C2 and C3, of immunoglobulin molecules It constitutes a Y-shaped basic area. Furthermore, the H chain separates C1 and C2 from the rest of the chain. It has a separate hinge region. The hinge region provides flexibility to four proteins are doing.

In a preferred embodiment, the protein complex of the present invention is partially or completely immunized against human immunization. It has a Y-shaped basic region that is the same as the C region of globulin. In this way, human immunoglobulin With the use of brine, the modified immunoglobulin is recognized as foreign by itself. This eliminates the problem of deterioration, making it easier to apply the method to human therapy. In addition, this The basic regions of the molecule provide the molecule with stability, Fc receptor binding, protein A Effector functions such as binding, complex fixation, and placental permeability can be imparted.

Modified sequences based on immunoglobulin molecules ensure that the modifications do not cause immune rejection. However, such limitations are within the scope of the present invention.

The basic region contains a binding site that binds to and activates CTL, and a binding site that binds to antigen. A joining part has been added. Antibody specific for CD3, a 07 surface protein is the binding site for Similarly, other CTL surface proteins capable of CTL activation Also included within the scope of the present invention are specific antibodies for.

These binding sites are located at the amino-terminal portion of the immunoglobulin-like Y-shaped basic region. Fixed by peptide bonds.

Antigen recognition binding sites are selected to exhibit specificity for a particular target organism . For example, the binding site of an antibody specific for a target organism is the amino acid in the arm of the basic region. It is fixed at the distal end.

In a preferred embodiment of the invention, the antigen recognition binding site is one of the Y-shaped basic regions. The CTL-specific antibody binding site is fixed to the arm of the CTL by a peptide bond, and the Y-shaped It is fixed to the other arm of the basic region of the protein by a peptide bond.

The hybrid immunoglobulin according to the present invention is effective against a wide range of viral infections. Ru. The hybrid immunoglobulin of the present invention causes cell death upon infection of host cells. Treatment for cases of prior infection with a virus that expresses the viral coat protein. Particularly suitable for medical treatment. In most cases, viral coats are formed in cells in this way. When proteins are expressed, they are formed on the cell surface. such an example and The four-magglutinin protein complex of the influenza virus, the mouse leukocyte protein complex, Env protein, RouS sarcoma virus Tamaeb protein, Examples include, but are not limited to, HIV protein. to infected cells During the initial infection, the protein expressed by the cell recognizes the cell receptor protein and binds to it. It is often the same viral coat protein that initiates the infection. this thing The same applies to HIV.

Anti-idiotypic antibodies, which have internal forms of microbial antigens, act on the TCR of T cells. It is known to stimulate humoral and cellular antimicrobial immunity, as well as antibodies against It is.

Preferred specific examples for producing such novel antibodies include genetic manipulation, This involves directly incorporating the antigen's gene sequence into the antibody. One way is , through genetic engineering, some of the immunoglobulin molecules are transferred to B cells or T cells. Such antibodies can be produced by substituting sequences corresponding to recognized HIV antigenic determinants. to be accomplished.

To give an example of the present invention, influenza virus nucleoproteins recognized by T cells A white (NP) epitope replaces the C region of the heavy chain of the antibody. This construct is 5P2 10 Expressed in myeloma cell line. Such infected 5P210 cells are NP killed by nibtope-specific T cells.

In the example shown below, two expression vectors pSV2gpt-91A3VH- CI gG2b and pSV2neo-91A3L are both called 9183 Anti-arsenate antibody (ant1-arsenate antibody) eavy chain) and L chain (ljghtchain) genes are integrated. It is rare. pSV2gpt-91A3VH-C1gG2b is the channel shown in Figure 3. 1. :, C region of IgG2b inserted into Hindll restriction enzyme site (constar+t regton) gene and jcoRI restriction enzyme site It has a rearranged 5.5 kb VHDJ gene of the 9183 antibody inserted. 5.5 The kb fragment also contains the heavy chain Ig promoter and enhancer. pSV2 neo-9], A3L has rearranged VL, CL genes and EcoRI, Ba The necessary regulatory genes are inserted into the mHI restriction enzyme site. These vectors - was cotransfected into 5P210, a non-secreting bone tumor cell line. On) - it was found that functional 9183 antibody was produced.

The VH of this antibody is derived from the 1588 strain, and the C region (Dsegment) is Likely involved in the antigen binding site. These views suggest that this C region is exposed on the surface. It suggests that there is.

In fact, due to the hydrophilicity of 91A 3 V H, the D region is exposed on the surface. It is predicted that For these reasons, 91A3VHDJ has the NP epitope. 1g chimera was selected for construction. The goal of this research is to achieve nine goals, as shown in Figure 3. The D region consisting of amino acids is converted into an NP CTL epitope consisting of 15 amino acids. It is to replace.

This epitope corresponds to amino acid residues 147-181 in the NP of the PR8 virus. Correspondingly, it induces virus-specific CTL in Ba1b/c mice, but C5 It is known that it is not induced in 7BL/6 mice.

Molecules of the invention are produced by recombinant DNA technology or chemical cross-linking. suitable Fuse the molecule in the correct orientation, introduce the gene into a suitable host cell, and express the protein. Methods of expression and purification are known in the art, examples of which are provided below. DNA There are many sources for details on cloning methods. For example, sample ( Sambrook et al. “Molecular Cloning Experiment Manual (Molewlar) Cloning A Laboratory Manual) J Cold Sp Ring Harbor Laboratory Press (Cold Spring Harbor See Laboratory Press) NY (1989).

Once the fusion gene has been cloned, the gene can be transferred (transferred) into a suitable host. The encoded protein is expressed.

The cloned gene is first inserted into a suitable vector or It is transferred into cells as near DNA and recombined with the host genome. Appropriate expression vector Examples of agents include plasmids, viruses, retroviruses, etc. Not limited to these. The choice of which vector is appropriate depends on which host is used for protein expression. It depends in part on the choice of use. Suitable hosts include bacteria, mammals, Cell lines derived from mammals, animal individuals such as transgenic mice, and cells derived from insects Examples include, but are not limited to, systems. Until now, cell lines derived from insects Although not used to express immunoglobulins, compared to mammalian cell lines, Due to the difference in glycoproteins, it is thought that more effective proteins are produced. insect-derived thin The cell system has lower maintenance costs and higher protein production compared to mammalian cells, so it can produce large amounts of protein. It is better suited for white matter production.

Genes expressed in insect-derived cell lines do not contain exons, so insect-derived Prior to expression in cell lines, exons must be excised.

Excision is relatively easy to perform, e.g., site-specifically using oligonucleotides. directly by mutagenesis or indirectly by cDNA cloning can be done.

Genes can be transferred to the host by any method well known in the art. . For example, gene transfer methods include calcium chloride in bacteria and eukaryotes. (CaC12)-mediated transfer, calcium phosphate (CaPO4)-mediated transfer, Relow Viral infections, including latent viral infections, electroporation, liposomal media Examples include mediated DNA transfer, microinjection, etc. Not exclusively.

Any suitable method for purifying proteins produced by a host may be used to practice the invention. give Webb et al. “Produced in baculovirus-infected insect cells” Cell surface expression of human CD4 and its purification Expression and Purification of Human CD4 Produced in Baculovirus-infeC,t ed　In5ec↑ce11s)Proc,, Notl, Aead, Su, LI SA, 85 Near 731-7735 (1989); Moran et al. Analysis and crossover of V region genes of antibodies specific to antigens of various influenza viruses Reactive idiotype riabIe-Region Genes and 5hared Cross reactjve 1dlotypes.

r Antibodies 5 specific for Antigens r　Various　Infbuenza　Viruses) Vir, Ia+w uno1. ．． 1:1-12 (1987).

The present invention is useful in directing cell-mediated immune responses to viral infections. child Here, HIV-infected cells are cited as an example of the use of the present invention, but other diseases may also be used. It should be considered that the invention can also be applied to diseases and is covered by the scope of the present invention.

As with all drugs, the effective amount of antibodies in this invention must be determined experimentally. No. Factors to consider include the condition being treated and whether the antibody forms a complex with the toxin. the method of administration of the pharmaceutical composition (i.e., intravenous, intramuscular); Note, subcutaneous administration, etc.) and number of administrations. Such factors are known in the art. It is within the physician's ability to make these decisions without undue experimentation. It is enough.

The following examples illustrate, but do not limit, the invention.

Example 1 - DNA cloning The 27 nucleotides encoding the D region of IgG were excised to correspond to the NP epitope. The procedure for inserting 45 bases is summarized in FIG. All enzymes are Used according to Kerr's instructions. New England Biolabs (New) England Biolabs), Beverly, Masachi ). Unless otherwise stated, DNA cloning is performed using Maniatis (M (1982).

Using this method, the VH region D region of the 91A3 anti-arsenate antibody was isolated from any of the following. Replace with .

(a) Consensus sequence of B cell epitopes in the cysteine loop of gp120. this Although epitope sequences are diverse, 24 epitopes obtained from 241 cultured strains of HIV By inferring from the sequence of No. 5, a common sequence was established. This common sequence amino acid The amino acid sequence corresponds to residues 301-319 of gp120 and is shown below.

Arg-Lys-5er-1ie-His-1ie-Gly-Pro-Gly- Arg-Ala-Phe-Tyr-Thr-Thr-Gly-Glu-11e- 11e(b) T cells of gag residues 12-35 in HXB2 culture of HIV epitope.

Gl u-Leu-Asp-Arg-Trp-Gl u-Lys-11e-Ar g-Leu-Arg-Pro-Gly-Gly-Lys-Lys-Lys-Ty r-Lys-Leu-Lys-Hi s-11e-Va 1(c) HIV-1 T cell epitope of reverse transcriptase. Residues 325-349.

Ala-11e-Phe-Gin-5er-5er-Het-Thr-Lys- 1ie-Leu-Glu-Pro-Phe-Arg-Lys-Gin-Asn- Pr.

-Asp-11e-Va l-11e-Thr-GlnSimply put, Croni The 5.5k fragment was inserted into the EcoRI restriction enzyme site of the pUc19 plasmid. This was done by subcloning b-91A3V HDJ. 2 The unique restriction enzyme sites (Ncol and ApeI, 63 g base pairs apart) It became clear that it surrounded area D.

Primers P1 and P3 shown in Figure 4 are fully complementary to the corresponding strands.

However, P2 matches the complementary strand up to the 5' terminal nucleotide of the D region. . (Black area) The remaining 30 nucleotides (shaded area) are the NP epitope.

Primer P4 contains a nucleotide complementary to the strand up to the 5' terminal nucleotide of the D region. Has cleotide. The remaining unmatched nucleotides are the 30th nucleotide of the NP epitope. Corresponds to the base. Among the overlapping nucleotides between P2 and P4, there is a 5prI restriction. A restriction enzyme site was created.

Enzymatic DNA amplification method (polymerase chain reaction, Two fragments are created by PCR). In a series of reactions, the P3 protein is added to the plasmid. By annealing the primer and P4 primer, a 570 base pair Fragments are created. In other series of reactions, P1 primer and P2 primer were used as primers. By annealing to the sumid, a 32B base pair fragment is created. To NP Both fragments are digested with 5pel to excise the overlapping sequences. NP epitome By ligation (Iigatfon) of two fragments containing each half of the 870 base pairs are formed containing the paired NP epitopes in a frame.

The next step was to remove both the original PUC19-VHDJ91A3 and the 870 base pair fragment. This is the process of digesting with restriction enzymes NcoI and ApeI. The 656 base pair fragment was deleted. By integrating the NP epitope into the transformed plasmid, the NP epitope can be inserted in place of the D region. A vector with a coding region is obtained. Follow the manufacturer's instructions for this 5.5. Co-transfection was performed using the Gene Pulsar transfer device (Biora d)). Plasmid psv2gp t-91A3-VHNPJ-CIgG2b and pSV2neo-91A3L was co-transfected into the non-secreting myeloma cell line 5P210. , by selection using mycophenolic acid and dieneticin (G418) , 91A 3-N P chimeric antibody synthesis and secretion were confirmed.

5P210 was co-introduced with the heavy chain carrying the HIV epitope and the original chain, resulting in Make the sufufutoma. Antibodies produced from these transfectomas are , used to induce humoral or cellular anti-HIV immunity.

Example 2 - Activity of chimeric antibodies Clones of NP-specific cytotoxic T cells were immunized with PR8 influenza virus. Irradiation produced in infected Ba1b/c mice and coated with 5 μg of NPs. Propagated in vitro by pancreatic cells. Cytotoxicity analysis was performed using “Cr-labeled Target cells and NP-specific CTL were incubated at a ratio of 10=1 for 4 hours. Ta. Coating of target cells with NPs was performed by adding 5 μg of peptide to 10 cells and adding 3 After incubation for 0 min and washing, J, I + nunol by ITO et al. .

Met, 103:229 (1987), “Cr Labeled using.

NP peptide (TYQRTRALVRTGMDP) is recognized by H-2 in the presence of on the other hand, the peptide (IASNENMDMESSST S) is a T cell epitope recognized in the presence of H-2DI' antigen.

The results are shown in Tables 1 and 2. In other words, influenza virus Chimeric Ig with a pitope binds to rabbit anti-NP antibody, and arsenate (arsenate) It has lost its bond to te>. This plays an important role in the binding ability to carmate. This is because the region was replaced with a viral peptide.

Table 1 pSV2gpt-91A3gpt-91A3VH and pSV2neo-91A3L Immunochemical analysis of immunoglobulin produced by SP210 co-transfected with Characteristic Binding to arsenate was determined using microtiter plates coated with arsenate BSA or BSA alone. - Measure binding by adding 10 ng of antibody to plate and incubating. The antibody was confirmed using +25) rat anti-mouse antibody. anti-isotype antibodies Binding was performed by adding antibody Long onto plates coated with rat anti-mouse kmAB. The bound antibody was determined by incubation with 1251 goat anti-mouse I Confirmation was made using gG2b antibody.

Table 22 Binding properties of 91A3 chimeric immunoglobulin (cp coat) Binding to arsenate-BSA , according to the method previously described in Example 1. Binding to rabbit anti-NP antibody Microtiter coated with anti-NP antibody purified by Nity chromatography This is done by adding the supernatant of transphenthoma to a plate and incubating it. Determination was made using combined antibody + 25I goat anti-mouse IgG2b antibody.

NP-specific CTLs are able to kill 5P210 transferred into the chimeric IG gene. This means that the NP epitope is present on the cell surface as well as in virus-infected cells. This suggests that it is expressed on the surface.

The data in Table 3 (Part A) shows the following. CTL clones are co-created with NP. PH10 and X31 influenza viruses as well as p815 cells Infected p815 cells can be killed. Unrelated NP (C57BL/6 Ma is known to be recognized by mouse CTL in the presence of H-2Db). No significant cell death is seen in coated p815 cells. Part B of the table is a chimera NPs that express Ig genes or kill NP-coated 5P210 cells Demonstrates the ability of specific CTL. Expressing vHw, vLw or both genes No cell death was observed in the cells. However, 5P210 V) Ic Clear cell death was observed in the VLW) Lancehu Ephthoma.

Table 3 5P21 transferred with a plasmid carrying the VH-NP chimera gene (V1(c) Cell death by NP-specific CTL of 0 cells *ND-not done ) From these results, the following is clear. Influenza recognized by CTL Cells into which the chimeric immunoglobulin gene having the enzyme epitope is transferred are C Killed by TL.

Bahia 1no, (" CD4 j' also CD3 minutes snoring FIG, 2 ←-2959-→141 294-145142 139-120001FIG , 4 Continuation of front page (51) Int, C1, 5 identification code Office serial number // (C12P 2110 2 C12R1:91) (72) Inventor Aviv, Zagower 2 New York, USA 10 028 New York Apartment 7AE 85th Street 150 I

Claims (4)

[Claims] 1. (a) At least one of the two C regions (CH2 and CH3) and the hinge region and which are arranged in a relationship similar to the Y-shaped structure characteristic of human immunoglobulins. (b) basic regions connected via sulfide bonds, (b) groups linked via peptide bonds; The first binding site bound to this region is the hybrid immunoglobulin and human can form a bond between the cytotoxic T-lymph class and the cytotoxic T-lymph class. a binding site capable of activating (c) is not identical to the first binding site and is bound to the basic region by a peptide bond; A hybrid antibody consisting of a second binding site, 2. Claim 1, wherein the first binding site specifically binds to CD3. Hybrid immunoglobulin according to item 1. 3. The second binding site specifically binds to the human immunodeficiency virus coat protein. The hybrid immunoglobulin according to claim 1, characterized in that: 4. Viral epi that elicits humoral-mediated and cell-mediated antiviral immune responses Chimeric immunoglobulin with TOP.