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 Table of Contents  
ORIGINAL ARTICLE
Year : 2016  |  Volume : 29  |  Issue : 2  |  Page : 379-382

Arginase-1 enzyme in B-cell non-Hodgkin lymphoma


Department of Clinical Pathology, Faculty of Medicine, Menoufia University, Menoufia, Egypt

Date of Submission02-Sep-2013
Date of Acceptance12-Dec-2013
Date of Web Publication18-Oct-2016

Correspondence Address:
Hanem M Badwy
Department of Clinical Pathology, Faculty of Medicine, Menoufia University, Quesna, Menoufia, 32631
Egypt
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/1110-2098.192422

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  Abstract 

Objectives:
Assessment of arginase-1 enzyme in B-cell non-Hodgkin lymphoma (NHL).
Background:
Arginase-1 enzyme is involved in the mechanism of immune suppression in NHL, which affects both the treatment outcome and the patient survival.
Patients and methods:
This study was carried out on 42 NHL patients attending the Oncology Department, Menoufia University Hospitals. Of the 42 NHL cases, 16 were relapsed or refractory (eight refractory: five cases relapsed within 1 year, two cases relapsed after 3 years, and one case after 4 years) and 26 were newly diagnosed cases. Follow-up for 24 months was carried out for the new cases. Sixteen of them achieved remission, two cases were relapsed (one case relapsed after 6 months and the other case relapsed after 9 months), whereas eight cases were lost to follow-up. Twenty age-matched and sex-matched individuals were selected as controls. A peripheral blood sample was drawn, and arginase-1 enzyme was analyzed by the enzyme-linked immunosorbent assay method.
Results:
The plasma level of arginase-1 enzyme was significantly increased in cases compared with controls. It was also increased in stages IV and III compared with stages I and II. Arginase-1 has the highest ratio in DLBCL compared with follicular lymphoma, which is higher than other indolent lymphomas. After patients were followed up for 2 years, the statistical data showed a highly significant difference in arginase-1 between patients with relapsed disease and patients with disease remission.
Conclusion:
The arginase-1 pathway is a main mechanism in the immune suppression in NHL as arginase-1 is increased in NHL patients. It is also increased in advanced stages of the disease and in a more aggressive pathology and in refractory and relapsed disease.

Keywords: Arginase-1, B-cell non-Hodgkin lymphoma, diffuse large B cell lymphoma


How to cite this article:
Ali KA, Al-Basuni MA, Alftooh MA, Radwan WM, Badwy HM. Arginase-1 enzyme in B-cell non-Hodgkin lymphoma. Menoufia Med J 2016;29:379-82

How to cite this URL:
Ali KA, Al-Basuni MA, Alftooh MA, Radwan WM, Badwy HM. Arginase-1 enzyme in B-cell non-Hodgkin lymphoma. Menoufia Med J [serial online] 2016 [cited 2019 Nov 20];29:379-82. Available from: http://www.mmj.eg.net/text.asp?2016/29/2/379/192422


  Introduction Top


The majority of hematological cancers involve the lymphoid system, and the majority of the lymphoid system neoplasms involve the B-cell lineage [1]. Non-Hodgkin lymphoma (NHL) represents a heterogeneous group of lymphocytic disorders ranging in aggressiveness from very indolent cellular proliferation to highly aggressive and rapidly proliferative processes. Although it is the fifth most common cancer in Egypt [2], it remains poorly understood and is largely incurable [3].

Systemic immune suppression is often seen in cancer patients and is thought to contribute to patient morbidity through tumor-mediated immune evasion. Patients with compromised immune systems, such as those with HIV infection or those on immunosuppressive medications, are at an increased risk of developing NHL [4]. Polymorphisms in host germline immune genes have also been associated with the risk of developing NHL [5] as well as the survival in NHL patients [6]. The presence of host immune cells in the tumor microenvironment has also been correlated with the treatment outcome and survival [7]. It is noteworthy that the reduction in the absolute count of circulating lymphocytes has been identified as a poor prognostic factor for the overall survival in newly diagnosed NHL [8] and as a predictor of poor treatment response [9].

Although evidence for the role of immune suppression in tumor establishment and pathogenesis is unquestionable, the mechanisms and the cellular phenotype of systemic immune suppression in NHL patients remain to be fully characterized [0].

Immune suppression occurs through various mechanisms, which involve adaptive immune response, innate immunity, and other nonimmunological processes. The best recognized mechanism for immune suppression depends on arginine and iNOS pathways [1].

Arginase-1 enzyme converts L-arginine into urea and L-ornithine, and is important in the metabolism of the nonessential amino acid L-arginine. Myeloid-deprived suppressor cells (MDSCs) expressing arginase-1 deplete L-arginine from the microenvironment and limit its availability to T cells. T cells deprived of L-arginine are deficient in the CD3 chain of the T-cell receptor and are arrested in the G0–G1 phase of the cell cycle, resulting in inhibition of both their function and their proliferation [2].


  Patients and Methods Top


This study was carried out on 42 NHL patients, 14 male and 28 female, with an age range of 33–71 years, attending the Oncology Department, Menoufia University Hospitals. Of the 42 NHL cases, 16 were relapsed or refractory (eight refractory: five cases relapsed within 1 year, two cases relapsed after 3 years, and one case after 4 years) and 26 were newly diagnosed cases. A follow-up of 24 months was carried out for the new cases. Of them, 16 cases achieved remission, two cases were relapsed (one case relapsed after 6 months and the other case relapsed after 9 months), whereas eight cases were lost to follow-up. Twenty age-matched and sex-matched individuals were selected as controls.

Sampling

A peripheral blood sample was drawn and dispensed into K3-EDTA evacuated tubes and centrifuged within 15 min; plasma was separated for arginase-1 enzyme analysis by the enzyme-linked immunosorbent assay (ELISA) method.

Analytical methods

Arginase-1 enzyme analysis by the ELISA method.

  1. Principle of the test:Arginase-1 was detected in stored plasma samples using the double-antibody sandwich ELISA kit (Glory Science Co. Ltd., Del Rio, TX, United States).

    Samples were added to enzyme wells precoated with human arginase monoclonal antibody; arginase antibodies labeled with biotin and combined with streptavidin–HRP were added to form an immune complex; then incubation and washing were carried out to remove the uncombined enzyme. When chromogen solutions A and B were added, the color of the liquid changed to blue, and the color finally became yellow under the effect of the acid. The chroma of color and the concentrations of the human substance arginase of the sample were positively correlated.

  2. Summary of the procedure:

    1. Reagents, samples, and standards were prepared.
    2. Samples were injected as follows:

      1. Blank well: only chromogen solutions A and B and stop solution were added; other operations were the same.
      2. Standard wells: standards and streptavidin–HRP were added (as the standard has already combined biotin antibody, it was not necessary to add the antibody).
      3. Test wells: samples were added and then both arginase–antibody and streptavidin–HRP were added.


    3. Incubation for 60 min at 37°C was performed.
    4. Plates were washed five times, chromogen solutions A, B were added and allowed to react for 10 min at 37°C.
    5. Stop solution was added, and the optical density value was measured within 10 min.


  3. Calculation of results:

    1. Optical density values of all the wells with the standard and the wells with the samples to be assayed were calculated.
    2. The standard curve diagram was made with the concentration of the standard from low to high and from left to right on the abscissa (X) axis and optical density values of the wells at 450 nm on the ordinate (Y) axis.
    3. Determination of the corresponding concentration range of each sample to be assayed on the standard curve diagram according to their optical density values.





  Results Top


Comparison between cases and controls

Comparison between cases and controls regarding the arginase-1 level showed a highly significant difference between them (case 170.9 ± 76.8, n = 43; control 96.2 ± 15.4, n = 20; P<0.0001).

Comparing the results of arginase-1 level in different stages

The percentage of arginase-1 was highly increased in disseminated disease (stages III and IV) than in limited disease (stage II 141.9 ± 70.5, n = 26; stages III and IV 218.1 ± 63.1, n = 16; P = 0.001).

Difference in arginase-1 among different pathologies

On comparing different pathological types of lymphomas, the highest ratio of arginase-1 was found in DLBCL, followed by follicular lymphoma (grade III), which was higher than other indolent lymphomas (follicular grade I and grade II, small-cell lymphoma and mucosa associated lymphoid tumor lymphoma) (diffuse 201.4 ± 71.6, n = 21; follicular 151.7 ± 77.9, n = 16; indolent 104.6 ± 13.9, n = 5; P = 0.014).

The relation between arginase-1 and treatment response ( first line)

A highly significant difference in the percentage of arginase-1 level was found between relapsed/refractory patients and in those who achieved remission during the follow-up period (relapsing 192.8 ± 66.2, n = 18; remission 127.2 ± 49.9, n = 16; P = 0.003).


  Discussion Top


NHL represents a heterogeneous group of lymphocytic disorders ranging in aggressiveness from very indolent cellular proliferation to highly aggressive and rapidly proliferative processes [3].

Systemic immune suppression is often seen in cancer patients and is thought to contribute to patient morbidity through tumor-mediated immune evasion. Patients with compromised immune systems, such as those with infection or on immunosuppressive medications, are at increased risk of developing NHL [4].

In recent years, MDSCs were recognized as important suppressors of immune response [3].

MDSCs inhibit immune responses through various mechanisms, which involve adaptive immune response, innate immunity, and other nonimmunological processes. The best recognized mechanism for immune suppression depends on arginine and iNOS pathways [1].

Arginase-1 converts L-arginine into urea and L-ornithine, and is important in the metabolism of the nonessential amino acidL-arginine. MDSCs expressing arginase-1 deplete L-arginine from the microenvironment and limit its availability to T cells. T cells deprived of L-arginine are deficient in the CD3 chain of the T-cell receptor and are arrested in the G0–G1 phase of the cell cycle, resulting in inhibition of both their function and proliferation [2].

Plasma levels of arginase-1 enzyme were elevated in NHL patients compared with controls.

Plasma levels of arginase-1 were elevated in higher stages of the disease, stages III and IV, and in more aggressive pathologies.

An elevated level of arginase-1 was found in relapsed disease or in cases refractory to treatment.

The result of this study agrees with Lin and colleagues, who studied CD14+ HLA-DR low/- monocytes in B-cell NHL and studied arginase-1 enzyme as a mechanism of their immunosuppressive effect, and stated the same results ([Table 1],[Table 2],[Table 3],[Table 4],[Table 5]).
Table 1: Descriptive clinical and laboratory data of all cases

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Table 2: Comparison between cases and controls regarding the percentage of arginase-1 level

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Table 3: Difference in arginase-1 at different stages

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Table 4: Difference of arginase-1 in different pathologies

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Table 5: Relation between arginase-1 and the treatment response (first line)

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  Conclusion Top


  1. Arginase-1 is increased in NHL patients.
  2. Arginase-1 is increased in higher stages of disease and in a more aggressive pathology.
  3. Arginase-1 is increased in refractory and relapsed disease.
  4. Arginase-1 may be involved in the mechanism of immunosuppression in B-cell NHL patients.


Conflicts of interest

There are no conflicts of interest.[13]

 
  References Top

1.
Dadi S, Le Noir S, Asnafi V, Beldjord K, et al. Normal and pathological V(D)J recombination: contribution to the understanding of human lymphoid malignancies. Adv Exp Med Biol 2009; 650:180–194.  Back to cited text no. 1
    
2.
Abdel-Fattah MM, Yassine OG. Non-Hodgkin's lymphomas in Alexandria, Egypt; incidence rates and trend study (1995–2004). Eur J Cancer Prev 2007; 16:479–485.  Back to cited text no. 2
    
3.
Han X, Li Y, Huang J, Zhang Y, Holford T, Lan Q, et al. Identification of predictive pathways for non-Hodgkin lymphoma prognosis. Cancer Inform 2010; 9:281–292.  Back to cited text no. 3
    
4.
Bhaskaran K, Brettle R, Porter K, et al. Systemic non-Hodgkin lymphoma in individuals with known dates of HIV seroconversion: incidence and predictors. AIDS 2004; 18:673–681.  Back to cited text no. 4
    
5.
Cerhan JR, Liu-Mares W, Fredericksen ZS, et al. Genetic variation in tumor necrosis factor and the nuclear factor-kappaB canonical pathway and risk of non-Hodgkin's lymphoma. Cancer Epidemiol Biomarkers Prev 2008; 17:3161–3169.  Back to cited text no. 5
    
6.
Habermann TM, Wang SS, Maurer MJ, et al. Host immune gene polymorphisms in combination with clinical and demographic factors predict late survival in diffuse large B-cell lymphoma patients in the pre-rituximab era. Blood 2008; 112:2694–2702.  Back to cited text no. 6
    
7.
Lenz G, Wright G, Dave SS, et al. Stromal gene signatures in large-B-cell lymphomas. N Engl J Med 2008; 359:2313–2323.  Back to cited text no. 7
    
8.
Talaulikar D, Choudhury A, Shadbolt B, Brown M. Lymphocytopenia as a prognostic marker for diffuse large B cell lymphomas. Leuk Lymphoma 2008; 49:959–964.  Back to cited text no. 8
    
9.
Oki Y, Yamamoto K, Kato H, et al. Low absolute lymphocyte count is a poor prognostic marker in patients with diffuse large B-cell lymphoma and suggests patients' survival benefit from rituximab. Eur J Haematol 2008; 81:448–453.  Back to cited text no. 9
    
10.
Lin Y, Gustafson MP, Bulur PA, Gastineau DA, Witzig TE, Dietz AB. Immunosuppressive CD14+ HLA-DR (low)/- monocytes in B-cell non-Hodgkin lymphoma. Blood 2011; 117:872–881.  Back to cited text no. 10
    
11.
Tamar T, Dina A, Aaron PK. Myeloid-derived suppressor cells – their role in haematooncological malignancies and other cancers and possible implications for therapy. Br J Haematol 2011; 153:557–567.  Back to cited text no. 11
    
12.
Ostrand-Rosenberg S, Sinha P. Myeloid-derived suppressor cells: linking inflammation and cancer. J Immunol 2009; 182:4499–4506.  Back to cited text no. 12
    
13.
Poschke I, Mougiakakos D, Hansson J, Masucci, GV, Kiessling R. Immature immunosuppressive CD14+ HLA-DR -/low cells in melanoma patients are Stat3hi and overexpress CD80, CD83, and DC-sign. Cancer Res 2010; 70:4335–4345.  Back to cited text no. 13
    



 
 
    Tables

  [Table 1], [Table 2], [Table 3], [Table 4], [Table 5]



 

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