Journal of Immunology and Clinical Research

Peripheral Blood Derived Treg Cells could Serve as Prognostic Marker of the Metastatic Cancer where TIL Derived Tregs are Not Accessible or Available for Analysis

Perspective | Open Access

  • 1. Department of Medicine, University of Connecticut, USA
+ Show More - Show Less
Corresponding Authors
Nitya G. Chakraborty, Department of Medicine, University of Connecticut, Farmington, CT 06030-3229, USA

•    Treg Cells
•    Metastatic Cancer
•    Natural Killer cells
•    Natural Killer T Cell


Although generating a robust and long-lived cytotoxic T lymphocyte (CTL) response to tumor associated antigens faces many constraints, adoptive immunotherapy has impressive clinical responses. The negative role of regulatory T cells (Treg) on immune responses, especially against “self” antigens (tumor associated antigens), has turned out to be a major constraint. In a recent report with tumor infiltrating lymphocyte (TIL) derived Treg cells fromhuman breast cancer and colon cancer, De simone’s group has clearly demostrated a poor prognosis of patients, associated wth increased number of functional Tregs from TIL. In this report we discuss, in human prostate cancer, that peripherally induced Treg (pTreg) cellsplay a major role in the down regulation of effective immunity against cancer whether those effectors are from innate immune system (NKT or NK cells) or from adaptive immunity via antigen specific cytotoxic T lymphocytes (CTL). Increase of peripheral blood (PBL) derived Treg cell number is directly related to higher gleason scores. PBL derived T cells, natural killer T cells (NKT) and natural killer cell (NK) showed significantly poor activity. Treg cells isolated from the patients with higher gleason score also showed increased suppressive activity in proliferation of conv CD4+ T cells.


Chakraborty NG (2017) Peripheral Blood Derived Treg Cells could Serve as Prognostic Marker of the Metastatic Cancer where TIL Derived Tregs are Not Accessible/ Available for Analysis. J Immunol Clin Res 4(2): 1044.


Recently De Simone et al. [1], demonstrated that tumorinfiltrating Treg cells display specific gene signatures that were also validated at the single-cell level. They have shown that tumorinfiltrating Treg cells were highly suppressive, up-regulated several immune-checkpoints, and expressed on the cell surfaces specific signature molecules. Their findings provided insights into the phenotypic, molecular and functional characteristics of human tumor-infiltrating Treg cells and define potential targets to improve tumor immunotherapy.

Metastatic diseases remain a serious challenge in cancer therapy. Regulatory T lymphocytes (Treg) appear to be a significant inducer of tolerance [2-7]. Here we discuss in human prostate cancer, that Treg cells play a major role in down regulation of effective immunity against cancers whether those effectors are from innate immune system (NKT or NK cells) or from adaptive immunity via antigen specific cytotoxic T lymphocytes (CTL) [2-9]. Lack of an effective way to engage CD4+ T cells as helper cells as well as effector cells in mitigating T reg cell activities has turned out to be the major problem. Dr. Gershon’s group was the first to describe T suppressor cells in cancer [2]. Then Dr. North’s group demonstrated that CD4+ T cells could function as suppressor cells in an animal tumor model [3]. We extended that basic observations and proved in clonal level that CD4+ T cells could function as suppressor cells in the human melanoma model [4,5]. These observations led to a substantial amount of work by others [6-9]. Dr. Sakaguchi’s group revived the topic of suppression by showing that a set of T cells emerging from the thymus naturally as CD4+ CD25+ T cells (hence called natural Treg (nTreg now called tTreg) has a critical role in the control of autoimmune pathology [6]. It is now clear that regulatory T cells are also generated in the periphery and are now referred to as p Treg cells [10].



Since Treg (CD4+ CD25+ Foxp3) cells play important role in down regulating tumor specific CTL response, we compared the Gleason score (prognostic marker) of prostate cancer patients’ disease with the number of Treg cells present in PBL of the respective patient. We found that the patients showing higher Gleason score also showed higher number of Treg cells in PBL as shown in Figure 1. When compared with of CD8+ T cell reactivity against prostate antigens, we observed that the patients having Gleason score from 5 or less had existence of antigen reactive CD8+ cells (CTL) and the reactivity of those cells was detectable for a prolonged period (checked two to three times in a period of two years). Whereas the CD8+ cells taken from the patients who had Gleason score 6 to 10, did show some initial reactivity against PSA peptides but that reactivity was lost as the number of functional T reg cells increased in PBL over the time of 2 years [11].


In innate immune system NK cells and NKT cells play very important role in anti tumor response. One interesting observation by Carnaud C, et al. (Cutting edge article in J Immunol) [12], where they showed that NKT cells rapidly activate NK cells in mouse and in human system. With our limited scope we wanted to see whether NKT cells could activate autologous NK cells in their effector phase also. We isolated NK cells by positively selecting CD56+ cells by using beads from Dynal (OSLO, Norway), from patients’ and normal donors’ PBL. We obtained an approximately 90% pure population of NK cells. We then separated the CD3+ CD56+ cells by positive selection with CD3+ dynal beads. We tested these two types of cells in NK killing assay using natural killer cell sensitive targets. Table 1 shows that the NKT cells do not show any significant killing of NK targets while CD56+ and CD3- NK cells showed significant killing. In a separate cytotoxicity assay we did mix NKT(CD56+ CD3+ cells) with (CD56+ CD3-) NK cells at the effector phase. The percent cytolytic activity of CD56+ NK cells was found to be increased significantly by the addition of double positive cells (Table 2) even when as low as ratio of 100 NK: 1 NKT cells was used. It appears that NKT cells might also enhance the activity of NK cells even at the effector phase. When the NK cells were taken out from a patient with higher Gleason score and higher number of T reg cells in PBL we detected significantly lower NK cell activity and no help from the NKT cells separated from the same patient (Table 1,2). The patient we tested here had a very high G-score and also very high Treg cell number. This observation with additional patients and normal donors and further comparing that with total number of T reg cells in PBL should be helpful to determine the prognosis and possible immunotherapeutic intervention for metastaic solid tumors.


Figure 2 shows a standard Treg assay in a mix culture of T regs and CD4+ CD25- T cells (Tconv) as responder cells upon stimulation with anti CD3 antibody with or without autologous dendritic cells.

T and B lymphocytes could specifically recognize tumor cells [13]. T lymphocytes recognizing tumor associated antigenderived peptides presented by major histocompatibility complex (MHC) molecules play a central rolein immunotherapy [14]. In fact, anti-tumor CD8+ T cell responsesmight arise automatically in cancer patients but those are disabled by varying conditions in the tumor microenvironment and tumor progresses [15]. The immunosuppressive mechanisms depend on the integrated action of in filtrating leukocytes, lymphocytes and the tumor that upregulate a range of modulatory molecules,collectively called immune checkpoints [16]. Therefore, thesearch for antagonistsof inhibitory molecules isprimary goal of current anti-tumor research [17]. In several clinical trials with CAR T cells being tested with reasonable success to overcome all the above mentioned constraints [18]. Treg cells are found to express very high level of these inhibitory molecules and thenumber of these Treg cells increases also in circulation as the disease progresses. In this regard we argue that tumor tissue might not be accessible by the surgeons to get TIL derived Tregs but PBL derived Tregs could serve as prognostic marker and could be usefull for doing some simple experiments for identifying target molecules for the improvement of therapeutic outcome.

Table 1: Comparison of killing of NK sensitive target by NK cells and NKT cells.

Effector cells 
% Lysis at E:T
  By NK cells By NKT cells
5:1 2.5:1 5:1 2.5:1
Normal Donor 1 20 16 2 0
Normal Donor 2 24 21 3 1
Patient 4 0 0 0

Table 2: Increase of NK killing activity with donor 1 and pa. NK cells when mixed with NKT cells.

Effector cells Target lysis at E:T =2.5:1
Donor 1 NK 16
Donor 1 NK +NKT (100:1) 25
Donor 1 NK + NKT (10:1) 36
Patient NK 0
Patient NK + NKT (10:1) 0



1. De Simone, Alberto A, Grazisa R, Paola G, Valeria R, Claudia P, et al. Transcriptional Landscape of Human Tissue Lymphocytes Unveils Uniqueness of Tumor-Infiltrating T Regulatory Cells. Immunity. 2016; 45: 1135-1147.

2. Gershon RK, Kondo K. Infectious immunological tolerance. Immunology. 1971; 21: 903-914.

3. Mills D, North RJ. Expression of passively transferred immunity against an established tumor depends on generation of cytolytic T cells in recipient. Inhibition by suppressor T cells. J Exp Med. 1983; 157: 1448-1460.

4. Mukherji, Chakraborty NG, Sivanandham M, Nashed AL, Sporn JR, Ergin MT. Clonal analysis of cytotoxic and regulatory T cell responses against human melanoma. J Exp Med. 1989; 169: 1961-1976.

5. Chakraborty NG, Twardzik DR, Sivanandham M, Ergin MT, Hellstrom KE, Mukherji B. et al, Autologous melanoma-induced activation of regulatory T cells that suppress cytotoxic response. J Immunol. 1990; 145: 2359-2364.

6. Sakaguchi S, Sakaguchi N, Asano M, Itoh M, Toda M. Immunologic selftolerance maintained by activated T cells expressing IL-2 receptor alpha-chains (CD25). Breakdown of a single mechanism of selftolerance causes various autoimmune diseases. J Immunol. 1995; 155: 1151-1164.

7. Beyer M, Schultze JL. Regulatory T cells in cancer. Blood. 2006; 108: 804-811.

8. Maloy KJ, Powrie F. Regulatory T cells in the control of immune pathology. Nat Immunol. 2001; 2: 816-822.

9. Piccirillo CA, Thornton AM. Cornerstone of peripheral tolerance: naturally occurring CD4+CD25+ regulatory T cells. Trends Immunol. 2004; 25: 374-380.

10. Karimi S, Chattopadhyay S, Chakraborty NG. Manipulation of regulatory T cells and antigen-specific cytotoxic T lymphocyte-based tumour immunotherapy. Immunology. 2015; 144: 186-196.

11. Chakraborty NG, Robert LS, Shikhar M, Elizabeth L, Pamela T, Jonathan RS, et al. Recognition of PSA-derived peptide antigens by T cells from prostate cancer patients without any prior stimulation. Cancer Immunol Immunother. 2003; 52: 497-505.

12. Carnaud C, Lee D, Donnars O, Park SH, Beavis A, Koezuka Y, et al. Cutting edge: Cross-talk between cells of the innate immune system: NKT cells rapidly activate NK cells. J Immunol. 1999; 163: 4647-4650.

13. Mariam JH, Eirini T, Karl SP, Sergio AQ, Charles S. Curr Opin Pharmacol. 2013; 13: 497-503.

14. Pardoll DM. The blockade of immune checkpoints in cancer immunotherapy. Nat Rev Cancer. 2012; 12: 252-264.

15. Galluzzi L, Aitziber B, Oliver K, Laurence Z, Guido K. Immunological Effects of Conventional Chemotherapy and Targeted Anticancer Agents. Cancer Cell. 2015; 28: 690-714.

16. Munn DH, Bronte V. Immune suppressive mechanisms in the tumor microenvironment. Curr Opin Immunol. 2016; 39: 1-6.

17. Zitvogel L, Galluzzi L, Smyth MJ, Kroemer G. Mechanism of action of conventional and targeted anticancer therapies: reinstating immunosurveillance. Immunity. 2013; 39: 74-88.

18. Jackson HJ, Rafiq S, Brentjens RJ. Driving CAR T-cells forward. Nat Rev Clin Oncol. 2016; 13: 370-383

Received : 22 Aug 2017
Accepted : 11 Sep 2017
Published : 13 Sep 2017
Annals of Otolaryngology and Rhinology
ISSN : 2379-948X
Launched : 2014
JSM Schizophrenia
Launched : 2016
Journal of Nausea
Launched : 2020
JSM Internal Medicine
Launched : 2016
JSM Hepatitis
Launched : 2016
JSM Oro Facial Surgeries
ISSN : 2578-3211
Launched : 2016
Journal of Human Nutrition and Food Science
ISSN : 2333-6706
Launched : 2013
JSM Regenerative Medicine and Bioengineering
ISSN : 2379-0490
Launched : 2013
JSM Spine
ISSN : 2578-3181
Launched : 2016
Archives of Palliative Care
ISSN : 2573-1165
Launched : 2016
JSM Nutritional Disorders
ISSN : 2578-3203
Launched : 2017
Annals of Neurodegenerative Disorders
ISSN : 2476-2032
Launched : 2016
Journal of Fever
ISSN : 2641-7782
Launched : 2017
JSM Bone Marrow Research
ISSN : 2578-3351
Launched : 2016
JSM Mathematics and Statistics
ISSN : 2578-3173
Launched : 2014
Journal of Autoimmunity and Research
ISSN : 2573-1173
Launched : 2014
JSM Arthritis
ISSN : 2475-9155
Launched : 2016
JSM Head and Neck Cancer-Cases and Reviews
ISSN : 2573-1610
Launched : 2016
JSM General Surgery Cases and Images
ISSN : 2573-1564
Launched : 2016
JSM Anatomy and Physiology
ISSN : 2573-1262
Launched : 2016
JSM Dental Surgery
ISSN : 2573-1548
Launched : 2016
Annals of Emergency Surgery
ISSN : 2573-1017
Launched : 2016
Annals of Mens Health and Wellness
ISSN : 2641-7707
Launched : 2017
Journal of Preventive Medicine and Health Care
ISSN : 2576-0084
Launched : 2018
Journal of Chronic Diseases and Management
ISSN : 2573-1300
Launched : 2016
Annals of Vaccines and Immunization
ISSN : 2378-9379
Launched : 2014
JSM Heart Surgery Cases and Images
ISSN : 2578-3157
Launched : 2016
Annals of Reproductive Medicine and Treatment
ISSN : 2573-1092
Launched : 2016
JSM Brain Science
ISSN : 2573-1289
Launched : 2016
JSM Biomarkers
ISSN : 2578-3815
Launched : 2014
JSM Biology
ISSN : 2475-9392
Launched : 2016
Archives of Stem Cell and Research
ISSN : 2578-3580
Launched : 2014
Annals of Clinical and Medical Microbiology
ISSN : 2578-3629
Launched : 2014
JSM Pediatric Surgery
ISSN : 2578-3149
Launched : 2017
Journal of Memory Disorder and Rehabilitation
ISSN : 2578-319X
Launched : 2016
JSM Tropical Medicine and Research
ISSN : 2578-3165
Launched : 2016
JSM Head and Face Medicine
ISSN : 2578-3793
Launched : 2016
JSM Cardiothoracic Surgery
ISSN : 2573-1297
Launched : 2016
JSM Bone and Joint Diseases
ISSN : 2578-3351
Launched : 2017
JSM Bioavailability and Bioequivalence
ISSN : 2641-7812
Launched : 2017
JSM Atherosclerosis
ISSN : 2573-1270
Launched : 2016
Journal of Genitourinary Disorders
ISSN : 2641-7790
Launched : 2017
Journal of Fractures and Sprains
ISSN : 2578-3831
Launched : 2016
Journal of Autism and Epilepsy
ISSN : 2641-7774
Launched : 2016
Annals of Marine Biology and Research
ISSN : 2573-105X
Launched : 2014
JSM Health Education & Primary Health Care
ISSN : 2578-3777
Launched : 2016
JSM Communication Disorders
ISSN : 2578-3807
Launched : 2016
Annals of Musculoskeletal Disorders
ISSN : 2578-3599
Launched : 2016
Annals of Virology and Research
ISSN : 2573-1122
Launched : 2014
JSM Renal Medicine
ISSN : 2573-1637
Launched : 2016
Journal of Muscle Health
ISSN : 2578-3823
Launched : 2016
JSM Genetics and Genomics
ISSN : 2334-1823
Launched : 2013
JSM Anxiety and Depression
ISSN : 2475-9139
Launched : 2016
Clinical Journal of Heart Diseases
ISSN : 2641-7766
Launched : 2016
Annals of Medicinal Chemistry and Research
ISSN : 2378-9336
Launched : 2014
JSM Pain and Management
ISSN : 2578-3378
Launched : 2016
JSM Women's Health
ISSN : 2578-3696
Launched : 2016
Clinical Research in HIV or AIDS
ISSN : 2374-0094
Launched : 2013
Journal of Endocrinology, Diabetes and Obesity
ISSN : 2333-6692
Launched : 2013
Journal of Substance Abuse and Alcoholism
ISSN : 2373-9363
Launched : 2013
JSM Neurosurgery and Spine
ISSN : 2373-9479
Launched : 2013
Journal of Liver and Clinical Research
ISSN : 2379-0830
Launched : 2014
Journal of Drug Design and Research
ISSN : 2379-089X
Launched : 2014
JSM Clinical Oncology and Research
ISSN : 2373-938X
Launched : 2013
JSM Bioinformatics, Genomics and Proteomics
ISSN : 2576-1102
Launched : 2014
JSM Chemistry
ISSN : 2334-1831
Launched : 2013
Journal of Trauma and Care
ISSN : 2573-1246
Launched : 2014
JSM Surgical Oncology and Research
ISSN : 2578-3688
Launched : 2016
Annals of Food Processing and Preservation
ISSN : 2573-1033
Launched : 2016
Journal of Radiology and Radiation Therapy
ISSN : 2333-7095
Launched : 2013
JSM Physical Medicine and Rehabilitation
ISSN : 2578-3572
Launched : 2016
Annals of Clinical Pathology
ISSN : 2373-9282
Launched : 2013
Annals of Cardiovascular Diseases
ISSN : 2641-7731
Launched : 2016
Journal of Behavior
ISSN : 2576-0076
Launched : 2016
Annals of Clinical and Experimental Metabolism
ISSN : 2572-2492
Launched : 2016
Clinical Research in Infectious Diseases
ISSN : 2379-0636
Launched : 2013
JSM Microbiology
ISSN : 2333-6455
Launched : 2013
Journal of Urology and Research
ISSN : 2379-951X
Launched : 2014
Journal of Family Medicine and Community Health
ISSN : 2379-0547
Launched : 2013
Annals of Pregnancy and Care
ISSN : 2578-336X
Launched : 2017
JSM Cell and Developmental Biology
ISSN : 2379-061X
Launched : 2013
Annals of Aquaculture and Research
ISSN : 2379-0881
Launched : 2014
Clinical Research in Pulmonology
ISSN : 2333-6625
Launched : 2013
Annals of Forensic Research and Analysis
ISSN : 2378-9476
Launched : 2014
JSM Biochemistry and Molecular Biology
ISSN : 2333-7109
Launched : 2013
Annals of Breast Cancer Research
ISSN : 2641-7685
Launched : 2016
Annals of Gerontology and Geriatric Research
ISSN : 2378-9409
Launched : 2014
Journal of Sleep Medicine and Disorders
ISSN : 2379-0822
Launched : 2014
JSM Burns and Trauma
ISSN : 2475-9406
Launched : 2016
Chemical Engineering and Process Techniques
ISSN : 2333-6633
Launched : 2013
Annals of Clinical Cytology and Pathology
ISSN : 2475-9430
Launched : 2014
JSM Allergy and Asthma
ISSN : 2573-1254
Launched : 2016
Journal of Neurological Disorders and Stroke
ISSN : 2334-2307
Launched : 2013
Annals of Sports Medicine and Research
ISSN : 2379-0571
Launched : 2014
JSM Sexual Medicine
ISSN : 2578-3718
Launched : 2016
Annals of Vascular Medicine and Research
ISSN : 2378-9344
Launched : 2014
JSM Biotechnology and Biomedical Engineering
ISSN : 2333-7117
Launched : 2013
Journal of Hematology and Transfusion
ISSN : 2333-6684
Launched : 2013
JSM Environmental Science and Ecology
ISSN : 2333-7141
Launched : 2013
Journal of Cardiology and Clinical Research
ISSN : 2333-6676
Launched : 2013
JSM Nanotechnology and Nanomedicine
ISSN : 2334-1815
Launched : 2013
Journal of Ear, Nose and Throat Disorders
ISSN : 2475-9473
Launched : 2016
JSM Ophthalmology
ISSN : 2333-6447
Launched : 2013
Journal of Pharmacology and Clinical Toxicology
ISSN : 2333-7079
Launched : 2013
Annals of Psychiatry and Mental Health
ISSN : 2374-0124
Launched : 2013
Medical Journal of Obstetrics and Gynecology
ISSN : 2333-6439
Launched : 2013
Annals of Pediatrics and Child Health
ISSN : 2373-9312
Launched : 2013
JSM Clinical Pharmaceutics
ISSN : 2379-9498
Launched : 2014
JSM Foot and Ankle
ISSN : 2475-9112
Launched : 2016
JSM Alzheimer's Disease and Related Dementia
ISSN : 2378-9565
Launched : 2014
Journal of Addiction Medicine and Therapy
ISSN : 2333-665X
Launched : 2013
Journal of Veterinary Medicine and Research
ISSN : 2378-931X
Launched : 2013
Annals of Public Health and Research
ISSN : 2378-9328
Launched : 2014
Annals of Orthopedics and Rheumatology
ISSN : 2373-9290
Launched : 2013
Journal of Clinical Nephrology and Research
ISSN : 2379-0652
Launched : 2014
Annals of Community Medicine and Practice
ISSN : 2475-9465
Launched : 2014
Annals of Biometrics and Biostatistics
ISSN : 2374-0116
Launched : 2013
JSM Clinical Case Reports
ISSN : 2373-9819
Launched : 2013
Journal of Cancer Biology and Research
ISSN : 2373-9436
Launched : 2013
Journal of Surgery and Transplantation Science
ISSN : 2379-0911
Launched : 2013
Journal of Dermatology and Clinical Research
ISSN : 2373-9371
Launched : 2013
JSM Gastroenterology and Hepatology
ISSN : 2373-9487
Launched : 2013
Annals of Nursing and Practice
ISSN : 2379-9501
Launched : 2014
JSM Dentistry
ISSN : 2333-7133
Launched : 2013
Author Information X