TCR/BCR PROFILING
This present disclosure relates to systems, methods, and compositions useful for profiling T cell receptor (TCR) and B cell receptor (BCR) repertoire using next-generation sequencing (NGS) methods. The present disclosure also relates to systems and methods for diagnosing, treating, or predicting infection, disease, medical conditions, therapeutic outcome, or therapeutic efficacy based on the TCR/BCR profile data from a subject in need thereof.
1 . A method of determining the TCR/BCR profile of a patient who has a disease selected from an infectious disease, an autoimmune disease, and allergic disease, or graft versus host disease, the method comprising:
a) isolating RNA from a sample from the patient;
b) enriching the isolated RNA for TCR/BCR genes using a set of TCR/BCR hybrid-capture probes and enriching for a targeted whole transcriptome panel using a set of transcriptome hybrid-capture probes;
c) sequencing the enriched RNA of (b) to generate sequencing data; and
d) analyzing the sequencing data to determine the TCR/BCR profile of the patient, wherein the set of TCR/BCR hybrid-capture probes comprises a first pool comprising TCR constant region probes, a second pool comprising TCR non-constant region probes, a third pool comprising BCR constant region probes, a fourth pool comprising BCR non-constant region probes, and a fifth pool comprising transcriptome hybrid-capture probes.
2 . The method of claim 1 , wherein the ratio of the first pool, second pool, third pool, and fourth pool within the set is 1:2.5:100:100.
3 . The method of claim 1 , wherein the ratio of the first pool, second pool, third pool, fourth pool, and fifth pool within the set is 1:2.5:100:100:10.
4 . The method of claim 3 , wherein 2% or less of the reads in the sequencing data map to TCR/BCR genes.
5 . The method of claim 1 , wherein the sample is a blood sample.
6 . The method of claim 1 , further comprising comparing the patient's TCR/BCR profile to a positive control profile.
7 - 14 . (canceled)
15 . A method of identifying TCR/BCR non-constant region sequences that are enriched in a cohort of patients with an infectious disease, an autoimmune disease, an allergic disease, or graft versus host disease, the method comprising:
a) isolating RNA from a sample from each patient in the cohort;
b) enriching the isolated RNA for TCR/BCR genes using a set of TCR/BCR hybrid-capture probes and enriching for a targeted whole transcriptome panel using a set of transcriptome hybrid-capture probes;
c) determining the sequence of the RNA of (b) to generate sequencing data;
d) analyzing the sequencing data to determine the TCR/BCR profile of the patients in the cohort; and
e) identifying TCR/BCR non-constant region sequences that are enriched in the cohort as compared to a control group without the disease,
wherein the set of hybrid-capture probes comprises a first pool of TCR constant region probes, a second pool of TCR non-constant region probes, a third pool of BCR constant region probes, and a fourth pool of BCR non-constant region probes, and a fifth pool of transcriptome hybrid-capture probes.
16 . The method of claim 15 , wherein the ratio of the first pool, second pool, third pool, and fourth pool within the set is 1:2.5:100:100.
17 . The method of claim 15 , wherein the ratio of the first pool, second pool, third pool, fourth pool, and fifth pool within the set is 1:2.5:100:100:10.
18 . The method of claim 17 , wherein 2% or less of the reads in the sequencing data map to TCR/BCR genes.
19 . The method of claim 15 , wherein the sample is a blood sample.
20 . (canceled)
21 . The method of claim 1 , wherein the infectious disease is Acinetobacter infection, Actinomycosis, African sleeping sickness (African trypanosomiasis), AIDS (acquired immunodeficiency syndrome), Amoebiasis, Anaplasmosis, Angiostrongyliasis, Anisakiasis, Anthrax, Arcanobacterium haemolyticum infection, Argentine hemorrhagic fever, Ascariasis, Aspergillosis, Astrovirus infection, Babesiosis, Bacillus cereus infection, Bacterial meningitis, Bacterial pneumonia, Bacterial vaginosis, Bacteroides infection, Balantidiasis, Bartonellosis, Baylisascaris infection, BK virus infection, Black piedra , Blastocystosis, Blastomycosis, Bolivian hemorrhagic fever, Botulism (and Infant botulism), Brazilian hemorrhagic fever, Brucellosis, Bubonic plague, Burkholderia infection, Buruli ulcer, Calicivirus infection (Norovirus and Sapovirus), Campylobacteriosis, Candidiasis (Moniliasis; Thrush), Capillariasis, Carrion's disease, Cat-scratch disease, Cellulitis, Chagas disease (American trypanosomiasis), Chancroid, Chickenpox, Chikungunya, Chlamydia, Chlamydophila pneumoniae infection (Taiwan acute respiratory agent or TWAR), Cholera, Chromoblastomycosis, Chytridiomycosis, Clonorchiasis, Clostridium difficile colitis, Coccidioidomycosis, Colorado tick fever (CTF), Common cold (Acute viral rhinopharyngitis; Acute coryza), Coronavirus disease 2019 (COVID-19), Creutzfeldt-Jakob disease (CJD), Crimean-Congo hemorrhagic fever (CCHF), Cryptococcosis, Cryptosporidiosis, Cutaneous larva migrans (CLM), Cyclosporiasis, Cysticercosis, Cytomegalovirus infection, Dengue fever, Desmodesmus infection, Dientamoebiasis, Diphtheria, Diphyllobothriasis, Dracunculiasis, Ebola hemorrhagic fever, Echinococcosis, Ehrlichiosis, Enterobiasis (Pinworm infection), Enterococcus infection, Enterovirus infection, Epidemic typhus, Erythema infectiosum (Fifth disease), Exanthem subitum (Sixth disease), Fasciolasis, Fasciolopsiasis, Fatal familial insomnia (FFI), Filariasis, Food poisoning by Clostridium perfringens , Free-living amebic infection, Fusobacterium infection, Gas gangrene (Clostridial myonecrosis), Geotrichosis, Gerstmann-Sträussler-Scheinker syndrome (GSS), Giardiasis, Glanders, Gnathostomiasis, Gonorrhea, Granuloma inguinale (Donovanosis), Group A streptococcal infection, Group B streptococcal infection, Haemophilus influenzae infection, “Hand, foot and mouth disease (HFMD)”, Hantavirus Pulmonary Syndrome (HPS), Heartland virus disease, Helicobacter pylori infection, Hemolytic-uremic syndrome (HUS), Hemorrhagic fever with renal syndrome (HFRS), Hendra virus infection, Hepatitis A, Hepatitis B, Hepatitis C, Hepatitis D, Hepatitis E, Herpes simplex, Histoplasmosis, Hookworm infection, Human bocavirus infection, Human ewingii ehrlichiosis, Human granulocytic anaplasmosis (HGA), Human metapneumovirus infection, Human monocytic ehrlichiosis, Human papillomavirus (HPV) infection, Human parainfluenza virus infection, Hymenolepiasis, Epstein-Barr virus infectious mononucleosis (Mono), Influenza (flu), Isosporiasis, Kawasaki disease, Keratitis, Kingella kingae infection, Kuru, Lassa fever, Legionellosis (Legionnaires' disease), Pontiac fever, Leishmaniasis, Leprosy, Leptospirosis, Listeriosis, Lyme disease (Lyme borreliosis), Lymphatic filariasis (Elephantiasis), Lymphocytic choriomeningitis, Malaria, Marburg hemorrhagic fever (MHF), Measles, Middle East respiratory syndrome (MERS), Melioidosis (Whitmore's disease), Meningitis, Meningococcal disease, Metagonimiasis, Microsporidiosis, Molluscum contagiosum (MC), Monkeypox, Mumps, Murine typhus (Endemic typhus), Mycoplasma pneumonia, Mycoplasma genitalium infection, Mycetoma, Myiasis, Neonatal conjunctivitis (Ophthalmia neonatorum), Nipah virus infection, Norovirus, “(New) Variant Creutzfeldt-Jakob disease (vCJD, nvCJD)”, Nocardiosis, Onchocerciasis (River blindness), Opisthorchiasis, Paracoccidioidomycosis (South American blastomycosis), Paragonimiasis, Pasteurellosis, Pediculosis capitis (Head lice), Pediculosis corporis (Body lice), “Pediculosis pubis (pubic lice, crab lice)”, Pelvic inflammatory disease (PID), Pertussis (whooping cough), Plague, Pneumococcal infection, Pneumocystis pneumonia (PCP), Pneumonia, Poliomyelitis, Prevotella infection, Primary amoebic meningoencephalitis (PAM), Progressive multifocal leukoencephalopathy, Psittacosis, Q fever, Rabies, Relapsing fever, Respiratory syncytial virus infection, Rhinosporidiosis, Rhinovirus infection, Rickettsial infection, Rickettsialpox, Rift Valley fever (RVF), Rocky Mountain spotted fever (RMSF), Rotavirus infection, Rubella, Salmonellosis , Severe acute respiratory syndrome (SARS), Scabies, Scarlet fever, Schistosomiasis, Sepsis, Shigellosis (bacillary dysentery), Shingles (Herpes zoster), Smallpox (variola), Sporotrichosis, Staphylococcal food poisoning, Staphylococcal infection, Strongyloidiasis, Subacute sclerosing panencephalitis, Bejel, Syphilis, Yaws, Taeniasis, Tetanus (lockjaw), Tinea barbae (barber's itch), Tinea capitis (ringworm of the scalp), Tinea corporis (ringworm of the body), Tinea cruris (Jock itch), Tinea manum (ringworm of the hand), Tinea nigra, Tinea pedis (athlete's foot), Tinea unguium (onychomycosis), Tinea versicolor ( Pityriasis versicolor ), Toxic shock syndrome (TSS), Toxocariasis (ocular larva migrans (OLM)), Toxocariasis (visceral larva migrans (VLM)), Toxoplasmosis, Trachoma, Trichinosis, Trichomoniasis, Trichuriasis (whipworm infection), Tuberculosis, Tularemia, Typhoid fever, Typhus fever, Ureaplasma urealyticum infection, Valley fever, Venezuelan equine encephalitis, Venezuelan hemorrhagic fever, Vibrio vulnificus infection, Vibrio parahaemolyticus enteritis, Viral pneumonia, West Nile fever, White piedra (tinea blanca), Yersinia pseudotuberculosis infection, Yersiniosis, Yellow fever, Zeaspora, Zika fever, or Zygomycosis.
22 . The method of claim 1 , wherein the autoimmune disease is type 1 diabetes, rheumatoid arthritis, lupus, celiac disease, Sjögren's syndrome, multiple sclerosis, polymyalgia rheumatica, ankylosing spondylitis, alopecia areata, vasculitis, psoriasis, or temporal arteritis.
23 . The method of claim 1 , wherein the allergic disease is contact dermatitis, asthma, anaphylaxis, or non-IgE-mediated food allergies related to atopic dermatitis.
24 . The method of claim 1 , wherein the patient has undergone a transplantation.
25 . The method of claim 1 , further comprising treating the patient for the disease.
26 . The method of claim 1 , further comprising, at repeating steps (a)-(d) for the patient at a later time.