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| Year : 2014 | Volume
: 25
| Issue : 1 | Page : 41-44 |
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| Analysis of serum and salivary immunoglobulin M in patients with orofacial epithelial cancers |
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J Lasisi Taye1, Kolude Bamidele2, A Lasisi Olawale3, E Akang Effiong4
1 Department of Physiology and Oral Pathology, University of Ibadan/University College Hospital, Ibadan, Nigeria
2 Department of Oral Pathology, University of Ibadan/University College Hospital, Ibadan, Nigeria
3 Department of Otorhinolaryngology, University of Ibadan/University College Hospital, Ibadan, Nigeria
4 Department of Pathology, University of Ibadan/University College Hospital, Ibadan, Nigeria
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| Date of Submission | 04-Mar-2013 |
| Date of Decision | 26-Mar-2013 |
| Date of Acceptance | 19-Apr-2013 |
| Date of Web Publication | 21-Apr-2014 |
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 Abstract | | |
Context: The physiological changes in the humoral immune system of patients with orofacial epithelial cancers (OECs) are considered key factors in the pathogenesis, prognosis, and management of these individuals.
Aim: This study assessed the serum and salivary immunoglobulin M (IgM) levels in patients with OECs.
Settings and Designs: This is a cross-sectional study of the serum and salivary IgM profile among patients with OEC and healthy controls.
Materials and Methods: There were 78 subjects comprising 30 patients with untreated OEC, 18 patients with OEC receiving treatment and 30 healthy, age and gender matched individuals. The serum and salivary samples from the participants were analyzed for IgM using the enzyme linked immunosorbent assay technique.
Results: The mean value of serum IgM in OEC patients receiving treatment was significantly lower compared to untreated OEC patients and healthy controls (P = 0.01). However, the mean serum IgM among untreated OEC patients was not significantly different compared with healthy controls. In contrast, the salivary IgM level did not show any significant difference among the three groups (P = 0.06). Furthermore, there was no correlation between the serum and salivary levels of IgM among the subjects.
Conclusion: The findings from this study suggest that serum IgM levels in OEC patients receiving treatment might be good biomarker while salivary IgM may not be reliable as a marker in these individuals. Keywords: cancer, immunoglobulin M, oro-facial, saliva, serum
How to cite this article:
Taye J L, Bamidele K, Olawale A L, Effiong E A. Analysis of serum and salivary immunoglobulin M in patients with orofacial epithelial cancers. Indian J Dent Res 2014;25:41-4 |
How to cite this URL:
Taye J L, Bamidele K, Olawale A L, Effiong E A. Analysis of serum and salivary immunoglobulin M in patients with orofacial epithelial cancers. Indian J Dent Res [serial online] 2014 [cited 2023 Sep 23];25:41-4. Available from: https://www.ijdr.in/text.asp?2014/25/1/41/131053 |
The mucosal immune system, with its local mechanisms, appears to be independent from systemic immunity. It represents the first line of defense against the uptake of macromolecules and infectious agents in the gastrointestinal, respiratory, and genitourinary systems. [1] However, its relation to the development and control of neoplasia is not well-understood, despite the fact that most human malignancies derived from epithelial tissues appear at sites where the secretory immune system is vigorously functioning. [2]
The physiological changes in the humoral immune system especially of patients with orofacial carcinomas are considered as important factors in the pathogenesis, prognosis, and management of cancer patients. In addition, the role of the humoral immune system has been considered an interfering or blocking phenomenon in carcinogenesis and several cytotoxic and tumor-antigen related antibodies have been described. [3],[4] Various specific quantitative changes of serum proteins and immunoglobulins have been documented in patients with cancers of various sites. [5] Similarly, quantitative changes in the serum and salivary IgA and IgG in orofacial carcinoma patients have been reported by many authors; [6],[7],[8] however, that of immunoglobulin M (IgM) is sparsely documented.
| Materials and Methods | |  |
This study received ethical approval from the Institution Ethics committee (UI/UCH/EC/09/014). The study population included patients with orofacial epithelial cancer (OEC) attending the dental and radiotherapy clinics of the hospital and healthy control subjects. The clinical bio data of the participants was obtained through a self-administered proforma. In this study, OEC included malignant epithelial tumors affecting the lips, tongue, gingival, and floor of the mouth. Included in the study were 30 participants with OEC who had not commenced treatment, 18 participants with OEC who had treatment (chemoradiotherapy). OEC patients were consecutive patients with histologically diagnosed epithelial cancer. Furthermore, included were 30 age and sex matched healthy individuals. Patients with infected tumors were excluded.
Control participants had a face and mouth examination by the principal investigator in order to exclude suspicious mucosal lesions as well as acute and chronic periodontitis. All healthy control individuals had not received any medication 1 month prior to the study. None of the healthy controls had a history of any chronic disease, prior malignancy, immunodeficiency, and autoimmune disorders. All patients and controls gave informed consent.
Collection of saliva sample
Whole non-stimulated saliva was collected by asking participants to spit into a graduated universal bottle for a period of 10 min. This was immediately transferred to sterilized tubes and frozen. The samples were stored at −20 o C until the time for immunoglobulin measurement.
Collection of a blood sample
Blood was collected simultaneously from peripheral veins in the upper arm using a size 21G needle on a 5 ml disposal syringe after cleaning the area with methylated spirit swab. Samples were collected in Ethylene diamine tetraacetic acid (EDTA) bottles.
The samples were centrifuged for 15 min at 2000 rpm and plasma was separated from the cellular components using a plastic Pasteur pipette and stored at −20 o C prior to use. The plasma was used for the analysis.
Quantification of immunoglobulin classes
The samples stored in the freezer were thawed in a refrigerator for 18 h (to preserve the immunoglobulins) and then centrifuged at 8000 rpm for 15 min. Serum and salivary IgM was quantified using the enzyme linked immunosorbent assay method. [9]
Assay protocol
Dilution of samples
The assay for quantification of IgM in samples requires that each test sample be diluted before use. A dilution of 1/10,000 was used for serum and saliva samples.
All reagents were brought to room temperature before use. A total of 100 μL of 6 standards were dispensed in duplicate with pipette. Then, 100 μL of sample (in duplicate) were transferred into pre-designated wells with pipette. The micro titer plate was incubated at room temperature for 30 min. Following incubation, the contents of the wells were aspirated. Each well was completely filled with appropriately diluted Wash solution and aspirate. This was repeated 3 times. The wells were completely filled with wash buffer. The plate was inverted and the contents were poured out in a waste container. This was followed by sharply striking the wells on absorbent paper to remove residual buffer. This was repeated 3 times for a total of four washes. Then, 100 μL of appropriately diluted enzyme-antibody conjugate was transferred to each well using the pipette. This was incubated at room temperature for 30 min. The plate was kept covered in the dark and leveled during incubation. The wells were washed and blotted as previously described. A total of 100 μL of Tetramethylbenzidine (TMB) Substrate Solution was transferred into each well using the pipette. The wells were incubated in the dark at room temperature for precisely 10 min. After 10 min, 100 μL of stop solution was added to each well. The absorbance (450 nm) of the contents of each well was determined and the plate reader was calibrated.
Data analysis
The main outcome variables were the serum and salivary levels of IgM in patients with OEC and healthy controls. The data were initially explored using the version 16 of the Statistical Package for Social Sciences (SPSS16). For serum IgM levels, means ± standard deviation (SD) and ranges were calculated while for salivary IgM levels, logarithm values were used to calculate mean values because data were not normally distributed. Test of significance in comparing these variables was performed using the one-way analysis of variance test with Dunnett's T post hoc test as appropriate. Salivary and serum immunoglobulins were correlated using the Pearson's correlation test. The level of statistical significance was P < 0.05 for all analyses.
| Results | | |
The participants comprised 38 males and 40 females between the ages of 28 years and 85 years (mean, 53.5 years; SD, 13.6 years). This included 30 participants with OEC who had not commenced treatment, 18 participants with OEC who had commenced treatment (chemoradiotherapy) and 30 healthy control participants. The OEC participants and controls were comparable in age and sex as shown in [Table 1].
The mean serum IgM levels in the untreated OEC patients, treated OEC patients and healthy controls were 355.8 ± 117.1 mg/dl, 256.7 ± 146.3 mg/dl and 365.3 ± 84.1 mg/dl respectively [Table 2] while the salivary median levels of IgM were 1.13 ± 0.51 mg/dl, 0.7 ± 0.2 mg/dl and 1.04 ± 0.48 mg/dl respectively [Table 3]. The mean value of serum IgM in treated OEC patients was significantly lower compared to untreated OEC patients and healthy controls (P = 0.01). However, there was no significant difference in the median salivary levels of IgM among the three groups (P = 0.06). There was no significant correlation between the serum and salivary IgM levels in the three groups [Table 4].
| Discussion | | |
The main finding in this study is a lower serum IgM level in patients with OEC receiving treatment compared to untreated OEC patients and healthy controls. In addition, serum IgM level was not significantly different between untreated OEC patients and healthy controls. The reduced serum level of IgM in OEC subjects receiving treatment indicates compromised systemic immune functions in these individuals. [10] This reduction could be a reflection of malnutrition and immune suppression in these individuals especially secondary to the treatment (chemoradiation) being received. [11],[12]
This finding is consistent with that of Parveen et al. [13] who reported a significant reduction in serum IgM in oral cancer subjects being treated with radiation and chemotherapy compared to the healthy individuals. In this study, the subjects in the treated group also had a combination of radiotherapy and chemotherapy, which might have resulted in further immune suppression as evidenced by the reduction in their serum IgM levels. In addition, the reduction in the serum IgM levels in these individuals could be attributed to compromised oral physiological environment that might have caused impaired mastication leading to malnutrition with subsequent immune suppression. In contrast to our finding, Khanna et al. [14] found that the treatment modality (surgery and chemotherapy) does not seem to affect serum immunoglobulin levels. The variations in findings may be explained by the differences in the forms of treatment received by the subjects.
However, Parveen et al. [13] and Khanna et al. [14] reported significantly elevated levels of serum IgM in oral cancer patients compared to healthy controls.
This study suggests that salivary IgM values may not differ between untreated OEC patients, treated OEC subjects and healthy controls. This may be attributed to the low quantity of IgM in saliva. Brandtzaeg [15] showed that surface epithelium acts as a passive molecular sieve favouring transmission of IgG, whereas the secretary epithelium is provided with some selective transport mechanisms specific for IgA and IgM; thus, making the salivary level of IgM low.
The lack of correlation between serum and salivary IgM among the subjects may suggest that salivary level of IgM is independent of the serum level.
We conclude from our findings that serum level of IgM may be a useful marker in monitoring patients with OEC receiving treatment while salivary IgM in these individuals does not seem to be. However, long-term longitudinal studies with a larger population may be needed to validate these findings.
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Correspondence Address:
J Lasisi Taye
Department of Physiology and Oral Pathology, University of Ibadan/University College Hospital, Ibadan
Nigeria
 Source of Support: None, Conflict of Interest: None  | Check |
DOI: 10.4103/0970-9290.131053
[Table 1], [Table 2], [Table 3], [Table 4] |
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