CN-121987771-A - Immune carrier microsphere carrying individuation MHC-II binding polypeptide and vaccine preparation and application thereof

Abstract

The invention relates to the technical field of immune carriers, in particular to immune carrier microspheres loaded with individual MHC-II binding polypeptides, and vaccine preparation and application thereof, wherein the microspheres are of shell-core structures and comprise cores and shells; the core microsphere is loaded with an individualized MHC-II binding polypeptide, the MHC-II binding polypeptide sequence is obtained by prediction and screening based on HLA genotyping results, each HLA allele corresponds to at least one high-affinity MHC-II binding polypeptide, and the shell is dextran or other polymer coating. The invention has the advantages that T, B epitope is separated inside and outside the microsphere to inhibit immune competition, thus obtaining better immune effect. The antibody can avoid the side effect of cross reaction, reduce the diversity of carrier molecules and avoid the side effect of excessive activation of T cells, the Th1 and Th2 epitopes can be arranged in the microsphere, thus not only generating antibody immunity but also generating T cell immunity function, the particle size of the microsphere can be controlled, and the immune function can be exerted without adjuvant.

Inventors

• TAN LISONG

Assignees

• 上海微球生物科技有限公司

Dates

Publication Date

20260508

Application Date

20251219

Priority Date

20250509

Claims (8)

1. 1. An immunocompromised microsphere carrying an individualized MHC-II binding polypeptide, characterized in that, The microsphere is of a shell-core structure and comprises a core and a shell, wherein the core microsphere is loaded with 3-12 individualized MHC-II binding polypeptides, MHC-II binding polypeptide sequences are obtained through computer prediction screening based on HLA genotyping results of inoculators, each HLA allele corresponds to at least one high-affinity MHC-II binding polypeptide, the shell is a glucan with a sulfhydryl group or other hydrophilic polymer coating layer, and a hydrophobic group of the shell is used as a coupling group for connecting antigens to construct vaccines, so that the T/B epitope microsphere is separated from the inside and the outside.
2. 2. The immunocompromised microsphere according to claim 1, wherein said immunocompromised microsphere comprises 3-12 high affinity MHC-II binding polypeptides selected in silico and at least one universal MHC-II binding polypeptide for assisting B cell production of antibodies, said universal MHC-II binding polypeptide being selected from at least one of the following: YFAVYLQETPY, AKFVAAWTLKAAA, QYIKANSKFIGITEL, FNNFTVSFWLRVPKVSASHLE, AWLEAQEEEEVGF, QYIKANSKFIGITELKK, EPRAPWIEQEGPEYWDQE, ADVEVYRAVTPLGPPD, DTLRSYYADWYQQKPG, FVNQHLAGSHLVEAL, LNEDLRSWTAADTAA。
3. 3. The immunocompromised microsphere comprising an individualized MHC-II binding polypeptide of claim 1, wherein said MHC-II binding polypeptide is obtained by affinity scoring screening according to HLA detection typing using IEDB ProPred, MHCPred, SVRMHC, ARB, SMM-align, netMHCIIpan, IEDB and NETMHCPAN predictive software.
4. 4. The immunocompromised microsphere bearing individualized MHC-II binding polypeptide of claim 1, wherein said microsphere has a particle size of 0.1-5 microns.
5. 5. A method of preparing an immunocompromised microsphere bearing an individualized MHC-II binding polypeptide according to claim 1, comprising the following three steps: coating MHC-II binding polypeptide self-assembled microspheres with dextran or other hydrophilic polymers; Coating biological microspheres loaded with the personalized MHC-II binding polypeptides with dextran or other hydrophilic polymers; mRNA microspheres expressing MHC-II binding polypeptides without signal peptide sequences.
6. 6. A vaccine prepared on the basis of any one of the immunocompromised microspheres 1-4 carrying the personalized MHC-II binding polypeptides, wherein the T/B epitopes are split inside and outside the microsphere, avoiding competitive inhibition of the T/B epitopes, which are located within the microsphere.
7. 7. The vaccine of claim 6, wherein Th2 epitope polypeptide and Th1 epitope polypeptide are loaded in the microsphere, and the vaccine is used for preparing T, B cell bifunctional vaccine with antibody immunity and cell immunity effects.
8. 8. Use of a vaccine loaded with individualized MHC-II binding polypeptide according to claims 6-7 for the preparation of a medicament for the treatment of tumors, cardiovascular diseases, hyperglycemic diseases, allergic diseases, autoimmune diseases, alzheimer's disease, viral infections, drug withdrawal, weight loss and other chronic diseases treatable with antibodies.

Description

Immune carrier microsphere carrying individuation MHC-II binding polypeptide and vaccine preparation and application thereof Technical Field The invention relates to the technical field of immune carriers, in particular to immune carrier microspheres loaded with individual MHC-II binding polypeptides and vaccine preparation and application thereof. Background 1. Therapeutic B cell vaccine research advances Monoclonal antibody technology was invented in the seventies of the last century, and this invention is undoubtedly a milestone in medical history that plays an irreplaceable role in disease detection and treatment. After this century, the technology of humanized monoclonal antibodies has matured, which has brought about a market trend for therapeutic antibody products, and more in clinical trials and preclinical research stages, which has shown that antibodies have increased the likelihood of disease treatment. The B cell vaccine can generate antibodies in human body for a long time, and has bright application prospect but few products which are successfully marketed in the aspect of treatment. The only antibody vaccine product currently on the market is CIMAvax-EGF, which is the only antibody vaccine product on the market in the field of tumor treatment. The vaccine is that an Epidermal Growth Factor (EGF) induces an organism to generate an EGF antibody, blocks the combination of EGF and a cancer cell surface receptor, and further inhibits the growth and proliferation of cancer cells. CIMAvax-EGF provides a new choice for adjuvant therapy of non-small cell lung cancer and brings hope to patients. The method is deeply significant in that the method provides a successful paradigm for the development of tumor therapeutic vaccines, proves the feasibility of resisting cancer by inducing organisms to produce specific antibodies, and indicates the direction for subsequent researches. In benign disease, there are many therapeutic monoclonal antibodies, therapeutic vaccines are mainly used for viral infection, most of which are in clinical trial phase. It is appreciated that therapeutic vaccines for hypertension have been put into phase III clinic over nearly 80 years of effort and are expected to be marketed several years later at home and abroad. This development is a possible upcoming new therapeutic modality for hypertensive patients, more as a milestone for therapeutic vaccines. The antibody plays a therapeutic role in three modes in vivo, namely, the product of the specific binding of the antigen-antibody is not directly discharged from the body, but is taken up and decomposed by phagocytes in the body. This process not only helps to clear the antigen-antibody complex, but also releases the antigenic peptide, further stimulating the T cell response and enhancing the immune system function (e.g., a neocrown vaccine). 2. Preventing binding of the functional molecule to the receptor and inhibiting its function (e.g., EGF vaccine). 3. Bind to antigens on tumor cell membranes and kill tumor cells (e.g., her2 vaccine) via ADCC effects. 2. Molecular immunology basis of B cell vaccine and carrier B cells produce antibodies, requiring dual signal activation. The first signal is the binding of an epitope (B epitope) to a B Cell Receptor (BCR), a process that determines the specificity of an antibody. The second signal is the cytokine that the Th epitope of the antigen induces T cell release. Exogenous antigen is internalized into B cells upon binding to B cell receptors. The antigen molecules are hydrolyzed into polypeptide fragments in lysosomes (Lysosome) and endosomes (Endosome) within the cell, and bind to MHC-II molecules of B cells, which are then displayed on the surface of the B cells. These MHC-II binding polypeptide epitopes (also known as Th epitopes or T epitopes) are presented to the T Cell Receptor (TCR) of cd4+ T cells. Subsequent release of cytokines activates B cells, determining the fate of B cell expansion and antibody production. MHC-II binding polypeptides and Th epitope peptides are one-time, referred to as MHC-II binding polypeptides for B cells or DC cells, and as T epitope polypeptides for T cells. In general, exogenous proteins contain both B and Th epitopes and antibodies can be raised when animals and humans are immunized. Therapeutic vaccines are mostly antigens of endogenous proteins or polypeptides. Since T cells capable of recognizing self (endogenous) antigens are clonally cleared during T cell development, the body lacks T cells capable of assisting in the production of antibodies to such antigens. Thus, there is a need to attach endogenous antigen molecules to exogenous immune carrier molecules (Immunocarrier) to provide exogenous Th epitopes that can be recognized by T cells, thereby breaking immune tolerance and producing antibodies. The foreign molecules conventionally used are OVA, BSA, KLH, phage or adenovirus, etc., which can provide sufficient diversity of MHC-II binding pol