20 participants are expected to be enrolled for the Phase Ib clinical trial，this trail is expected to be finished in 20 months.

This study is to investigate the safety and efficacy of tumor infiltrating lymphocyte (TIL) therapy in patients with advanced melanoma. Autologous TILs are expanded from tumor resections or biopsies and infused i.v. into the patient after NMA lymphodepletion treatment with hydroxychloroquine and cyclophosphamide.

Inclusion Criteria:

1. Signed the informed consent form (ICF) and able to comply with the visits and related procedures specified in the protocol;

2. Aged ≥18 years and ≤75 years, regardless of gender;

3. Patients with unresectable advanced, recurrent or metastatic melanoma (excluding uveal melanoma) who have failed standard treatment with PD-1 antibodies, etc. (if BRAF mutation is carried, BRAF and MEK inhibitor treatment failure);

4. TILs can be isolated from a surgically resectable tumor region: the tissue volume must be >150mm3, and the lesion has not received local treatment (such as radiotherapy, radiofrequency ablation, oncolytic virus, etc.) or progressed after local treatment;

5. There are still at least 1 measurable lesion (according to RECIST1.1 criteria [see Appendix 4]) even after TIL sampling and resection of surgically resectable tissue;

6. ECOG performance status 0-1;

7. Expected survival time >3 months;

8. With sufficient hematology and end-organ function as defined by the following laboratory test results, the test results must be completed and issued within 7 days before tumor tissue collection:

   • White Blood Cell (WBC)≥2.5×10^9/L#
   • Absolute Lymphocyte Count (ANC)≥1.5×10^9/L;
   • Absolute Lymphocyte Count(ALC)≥0.7×10^9/L;
   • Platelet≥100×10^9/L#
   • International Normalized Ratio#INR#≤1.5×ULN;
   • Activated Partial Thromboplastin Time#APTT#≤1.5×ULN;
   • Serum Creatinine (Scr)≤1.5mg/dL (or 132.6μmol/L) or Creatinine
   • Clearance≥60mL/min
   • Urinalysis: urine protein less than 2+, or 24-hour urine protein <1g;
   • Alanine aminotransferase(AST/SGOT) ≤3×ULN;
   • Alanine aminotransferase (ALT/SGPT) ≤3×ULN;
   • Total Bilirubin(TBIL)≤1.5×ULN#

9. * Premenopausal women who have not undergone sterilization surgery must agree to use effective contraception measures from the start of study treatment (preconditioning) to one year after cell infusion, and the serum pregnancy test during the screening period must be negative; *Men who have not undergone sterilization surgery must agree to use effective contraception measures from the start of study treatment (preconditioning) until one year after cell infusion;

10. No absolute or relative contraindications for surgery;

11. Any melanoma treatment methods, including radiotherapy, chemotherapy, endocrine therapy, targeted therapy, immunotherapy, tumor embolization, or traditional Chinese medicine/herbal medicine treatment with anti-tumor indications, must be stopped 28 days before infusion. If a small molecular targeted drug was used in the previous treatment, the withdrawal time can be shortened to 5 half-lives of the drug used;

12. Good compliance and able to adhere to the study visit plan and other agreement requirements.

Exclusion Criteria:

1. Participation in a clinical trial of another drug or biologic therapy or receipt of a comparable cellular therapy within 28 days prior to infusion;
2. Combination of 2 or more malignant tumors, except: Eradicated malignant tumors that have been inactive for ≥5 years prior to study entry and are at minimal risk of recurrence; adequately treated non-melanoma skin cancer or malignant nevus of freckle-like nevus without evidence of disease recurrence; adequately treated carcinoma in situ without evidence of disease recurrence;
3. Has received live attenuated vaccination after signing informed consent or is scheduled to receive it during the study;
4. Has not recovered from a prior procedure or treatment-related adverse reaction to ≤ grade 1 nci ctcae 5.0 (except for toxicities such as alopecia, etc., which in the judgment of the investigator pose no safety risk);
5. Known history of allergy to streptomycin, ciprofloxacin, or micafungin or allergy to any component of the infused product formulation;
6. Uncontrolled co-morbidities including, but not limited to, uncontrolled arterial hypertension (systolic blood pressure ≥160 mmhg and/or diastolic blood pressure ≥100 mmhg) even with standardized treatment or any unstable cardiovascular disease including transient ischemic attack, cerebrovascular accident, myocardial infarction, unstable angina pectoris within 6 months prior to enrollment; new york heart association ( nyha class iii or iv congestive heart failure with an ejection fraction <50%; or severe cardiac rhythm or conduction abnormalities, such as ventricular arrhythmias, degree ii-iii atrioventricular block, etc., requiring clinical intervention; ecg results showing clinically significant abnormalities or a qtcf ≥450ms (if the first test is abnormal, it may be retested at least 5 minutes apart twice and the combined result/mean value to determine eligibility) ;
7. Patients with esophageal or gastric varices that require immediate intervention (e.g., taping or sclerotherapy) or are considered to be at high risk for bleeding based on the opinion of the investigator or consultation with a gastroenterologist or hepatologist, have evidence of portal hypertension (including splenomegaly detected on imaging), or have a prior history of variceal bleeding must have undergone endoscopic evaluation within 3 months prior to enrollment;
8. Uncontrolled metabolic disorders, such as diabetes mellitus known to be uncontrolled, or other non-malignant organ or systemic diseases or secondary reactions to cancer, and which can lead to higher medical risk and/or uncertainty in survival evaluation;
9. Hepatic encephalopathy, hepatorenal syndrome or child-pugh class b or more severe cirrhosis, liver failure;
10. Comorbidity with other serious organic or psychiatric disease;
11. Have an active systemic infection requiring treatment with positive blood cultures or imaging evidence of infection, including but not limited to active tuberculosis;
12. Be hiv-positive, have a positive serologic test for syphilis, or have clinically active hepatitis a, b, or c, including viral carriers: Hepatitis b, excluding those who are HBsAg-positive; hepatitis c, excluding those who are HCVAb-positive;
13. Who have used, or in the judgment of the investigator have a co-morbid condition requiring the use of glucocorticosteroids or other immunosuppressive medications during the trial within 4 weeks prior to pretreatment, excluding topical glucocorticosteroids by nasal spray, inhalation, or other routes or physiologic doses of systemic glucocorticosteroids (i.e., no more than 10 mg/day of prednisone or equivalent dose of other glucocorticosteroids), or who have an active autoimmune disease ( eczema, vitiligo, psoriasis, alopecia areata, or grave's disease that does not require systemic treatment within the last 2 years, other autoimmune diseases that are not expected to recur, and hypothyroidism requiring only thyroid hormone replacement therapy, and subjects with type i diabetes mellitus requiring only insulin replacement therapy may be enrolled);
14. Any nci ctcae5.0 immune-related adverse effect (irae) grade ≥ 3 during any prior period of immunotherapy receipt;
15. History of organ allograft, allogeneic stem cell transplantation and renal replacement therapy;
16. Pulmonary fibrosis, interstitial lung disease (both past history and current), and acute lung disease;
17. Clinically uncontrollable third space effusions, such as pleural and abdominal effusions that cannot be controlled by drainage or other means prior to enrollment;
18. Patients with known molluscum contagiosum metastases;
19. Patients with clinically symptomatic central nervous system metastases (e.g., cerebral edema, need for hormonal intervention, or progression of brain metastases). Patients with prior treatment for brain metastases, such as clinical stability (mri) that has been maintained for at least 2 months and who have discontinued systemic hormone therapy (dose >10 mg/day prednisone or other equipotent hormone) for >4 weeks may be included;
20. Women who are pregnant or breastfeeding;
21. History of allogeneic t-cell and nk-cell therapy;
22. If the investigator believes that other circumstances are not suitable for enrollment.