|Year : 2019 | Volume
| Issue : 1 | Page : 47-50
Helicobacter pylori in benign versus malignant laryngeal lesions
Mohammad W El-Anwar1, Ashraf Raafat2, Mohammad Abdehady3, Eman A Eissa4
1 Department of Otorhinolaryngology-Head and Neck Surgery, Zagazig University, Egypt
2 Department of Otorhinolaryngology, Alahrar Teaching Hospital, Zagazig, Egypt
3 Department of Otorhinolaryngology, Faculty of Medicine, Zagazig University, Egypt
4 Department of Clinical Pathology, Benha Teaching Hospital, Benha, Egypt
|Date of Submission||17-Apr-2018|
|Date of Acceptance||23-Oct-2018|
|Date of Web Publication||14-Feb-2019|
Department of Otorhinolaryngology-Head and Neck Surgery, Faculty of Medicine, Zagazig University, Zagazig, 12411
Source of Support: None, Conflict of Interest: None
Objective The aim was to investigate the existence of immunoglobulin (Ig)G antibody against Helicobacter pylori (H. pylori) in blood sample from patients having laryngeal lesions and comparing its level in benign versus malignant lesions.
Patients and methods Under general anesthesia, direct laryngoscopy was performed for patients having laryngeal lesions, and biopsy was taken and sent for histopathology. Anti-H. pylori IgG antibodies were measured by enzyme-linked immunosorbent assay from venous blood samples from each patient.
Results Within the included 56 patients, 30 had benign lesion and 26 (46%) had squamous cell carcinoma (SCC). Overall, 73.3% of patient with benign lesions were seropositive and 92.3% of patient with laryngeal SCC were seropositive. The mean anti-H. pylori IgG antibody level was significantly (P=0.0041) higher in patients who had SCC (23.93±19.6) than patients who had benign laryngeal lesions (38.9±27.5).
Conclusion Laryngeal lesions are commonly associated with H. Pylori infection and showed significantly higher level in laryngeal SCC than benign lesions, reflecting more association of larynx cancer with H. pylori infection.
Keywords: benign, H. pylori, larynx, malignant
|How to cite this article:|
El-Anwar MW, Raafat A, Abdehady M, Eissa EA. Helicobacter pylori in benign versus malignant laryngeal lesions. Egypt J Otolaryngol 2019;35:47-50
|How to cite this URL:|
El-Anwar MW, Raafat A, Abdehady M, Eissa EA. Helicobacter pylori in benign versus malignant laryngeal lesions. Egypt J Otolaryngol [serial online] 2019 [cited 2020 Jun 4];35:47-50. Available from: http://www.ejo.eg.net/text.asp?2019/35/1/47/251311
| Introduction|| |
Helicobacter pylori (H. pylori) are gram-negative microaerophilic microorganisms. It is estimated that more than 50% of the population worldwide carry this bacterium in their gastrointestinal tract . It has been verified that H. pylori plays an important role in the pathogenesis of peptic ulcers, chronic gastritis, gastric lymphoma, and adenocarcinoma . Immigration of H. pylori in the upper aerodigestive zone was proven , and the association between H. pylori and laryngeal malignancy was investigated; however, it is still a matter of controversy .
Laryngeal squamous cell carcinoma (SCC) is one of the most common malignant tumors of the head and neck, representing approximately 25% of the cases . The potential for epithelial and mucosal inflammation and damage could result in chronic harm and epithelial cell proliferation, leading to laryngeal pathology ,.
The aim of this study was to investigate the existence of immunoglobulin (Ig)G antibodies against H. pylori in blood sample from patients who had laryngeal lesions and to compare H. pylori level in benign versus malignant lesions.
| Patients and methods|| |
After approval by the institutional review board and obtaining a written informed consent from the patients, patients having laryngeal lesions who were scheduled for elective direct laryngoscopy for either biopsy or excisional biopsy in Otorhinolaryngology Department, Zagazig University Hospitals, and Benha teaching hospitals, during the period from February 2015 to April 2017 were chosen to participate in the study. Patients who had known hypersensitivity to used anesthetic drugs, unfit patients for general anesthesia, congenital laryngeal lesions, and syndromic patients were excluded from the study. Nonsteroidal anti-inflammatory drugs were avoided for 2 weeks before surgery. All patients were subjected to full history taking, general and local clinical examination, and routine preoperative laboratory testing.
Under general anesthesia, direct laryngoscopy was performed, and a biopsy was taken if the lesion was suspected to be malignant or total surgical excision for clinically suspected benign lesion, and all biopsies were sent for histopathology.
Quantitative determination of IgG antibody against H. pylori was done by enzyme-linked immunosorbent assay (ELISA).
Specimen collection and storage
Five ml of venous blood sample was taken from each patient and left to clot at 4°C in a sterile, clean, dry tube. After clotting, the samples were centrifuged for 10 min at 5000 rpm. Serum was separated and stored immediately at −20°C till the time of analysis.
Anti-H. pylori IgG antibodies were measured by ELISA using commercially available kit from RD Radio Diagnostics (catalog no. E HLG KO8, www.RDlab.com; Germany) Principle of the test: calibrators and unknowns are incubated in microtitration wells coated with purified and inactivated H. pylori antigen. After incubation and washing, the wells are treated with the conjugate, composed of anti-human IgG antibodies labeled with peroxidase. After a second incubation and washing step, the wells are incubated with the substrate tetramethylbenzidine. An acidic stopping solution is then added, and the degree of enzymatic turnover of the substrate is determined by wavelength absorbance measurement at 450 nm. The absorbance measured is directly proportional to the concentration of anti-H. pylori IgG antibodies present. Results: the cutoff control corresponds to calibrator 1. If the absorbance of the sample is higher than that of the cutoff, the sample was positive for the presence of specific IgG. The ratio between the average optical density of the sample and that of the cutoff was calculated. The sample was considered positive, if the ratio was more than 1.1; doubtful, if ±10% of the cutoff; and negative, if the ratio was less than 0.9. Results were expressed in u by interpolating the optical density values of the 5 calibrators and comparing the value of the sample with this curve.
Statistical analysis was performed using SPSS 14.0 statistical software for Windows (SPSS Inc., Chicago, Illinois, USA). The significance level was set at P less than or equal to 0.05.
| Results|| |
Within the included 56 patients, 30 (53.5%) patients were histopathologically proved to have benign lesion, including 24 (80%) males and six (20%) female. 20/30 (66.6%) cases were polyp, 6/30 (20%) were nodules, 2/30 (6.6%) was cyst and 2/30 (6.6%) was Reinke’s edema. On the contrary, malignant lesions were proved in 26 (46%) patients, and all were males and had SCC ([Table 1]).
|Table 1 Epidemiological and anti-H. pylori immunoglobulin G antibody results in benign and malignant laryngeal lesions|
Click here to view
The mean age of all patients was 47.57±11.9 (age range: 28–70) years. The mean age for patients with benign lesions was 40.8±9.88 years and in malignant lesions was 40.3±8.34 years. A total of 46 (82.1%) patients were seropositive, with 22/30 (73.3%) benign and 24/26 (92.3%) malignant, whereas 10 (17.85%) patients were seronegative, with eight (26.7%) benign and two (7.7%) malignant. Anti-H. pylori IgG antibodies level in patients who had benign lesions ranged from 3.2302 to 85.4 (mean=38.9±27.5) whereas anti-H. pylori IgG antibodies level in malignant lesions ranged from 8.247 to 87.275 (mean: 23.93±19.6). The mean anti-H. pylori IgG antibodies level was significantly (t=2.9967, P=0.0041) higher in patients who had malignant laryngeal lesions (23.93±19.6) than patients who had benign laryngeal lesions (38.9±27.5) ([Table 1]), reflecting significance association between H pylori and cancer larynx, which could be attributed to the importance of H. pylori level in pathogenesis of cancer larynx. We took into consideration that both groups of patients were matched for age; all patients were smokers and nonalocoholics.
| Discussion|| |
H. pylori infection, resulting from a gram-positive spiral-shaped or curved rod, is one of the commonest bacterial infections in human , with a prevalence reaching 90% in underdeveloped countries and 30% in developed countries .
Although the stomach is the primary site of this bacterium, H. pylori could also be detected in calculus, pharyngeal lymphoid tissue, saliva, nasal and sinus mucosa, oropharynx, tympanic cavity secretion, and larynx .
The mean age of our patients was 47.57±11.9 years near to the mean age reported by Barakat et al.  and Siupsinskiene et al. .
Malignant lesions were proved in 46% of included patients, which is near to the percent registered by Barakat et al.  However, Siupsinskiene et al.  found that 16.67% patients had malignant lesion. This is mostly because we included only cases scheduled for surgery, so all malignant lesions were included, whereas many patients with benign lesions do not prefer to undergo surgery.
The aim of this study was to determine the prevalence of H. pylori in the patients with benign and malignant laryngeal diseases.
ELISA is noninvasive serology method for detection of H. pylori. It is rapid test, and even though it is less sensitive, it is specific. This serological test is relatively widely available and cheap; therefore, it may be helpful in screening or in confirming the presence H. pylori infection . Although serology is not appropriate for determining the underlying gastrointestinal disease , his is not our concern in the current study. Thus, we used ELISA serology test for measuring anti-H. pylori IgG antibodies, which is available in our institute and could quantitatively measure antibody level. ELISA was used similar to Urita et al.  and Rezaii et al. .
Overall, 82.1% of our patients who had laryngeal pathology scheduled for surgery were seropositive. Thus, laryngeal lesions are commonly associated with H. pylori infection. In addition, 73.3% of patient with benign lesions were seropositive and 92.3% of patient with laryngeal SCC were seropositive, reflecting the more association of cancer larynx with H. pylori infection.
On quantitative measure of anti-H. pylori IgG antibodies level, the view becomes clearer as the mean anti-H. pylori IgG antibodies level was significantly (P=0.0041) higher in patients who had malignant laryngeal lesions than patients who had benign lesions, reflecting significance association between H. pylori and cancer larynx, which could be attributed to the importance of H. pylori level in etiopathogenesis of cancer larynx. We took into consideration that benign and malignant groups of patients were matched for age and sex. Importance of detection of anti-H. pylori IgG antibodies level with laryngeal lesions might direct attention to treatment of H. pylori infection in the postoperative regime for laryngeal pathology.
Barakat et al.  similarly commonly detected positive anti-H. pylori IgG antibody with laryngeal lesions but did not detect significant differences between benign and malignant lesions. Similarly, in 2008, Fang et al.  hypothesized that H. pylori may be an etiological factor of VF polyps.
The important association between H. pylori infection and the development of laryngeal cancer was proved by a meta-analysis by Tominaga , which indicated that the risk of developing laryngeal cancer is twice as high for the patients infected with H. pylori . Using serological studies, H. pylori infection was significantly more often exist in patients operated for malignant than benign laryngeal lesions, which suggests that H. pylori may be an independent factor of laryngeal SCC ,,,. Similarly, Rezaii et al. , using ELISA for detection of anti-H. pylori IgG antibodies, concluded that H. pylori is an independent risk factor for laryngohypopharyngeal carcinoma. Similar results were also reported by Titiz et al. , using PCR to detect genetic material from H. pylori in the larynx, and recorded significantly more H. pylori infection with cancer larynx than benign laryngeal diseases. In agreement with our results, some studies detected high prevalence of H. pylori in patients with laryngeal SCC by serology ,,,.In contrast, Nurgalieva et al. , using ELISA to detect antibodies to H. pylori, found that H. pylori infections do not either protect against or promote laryngopharyngeal carcinoma.
Compared with ELISA, PCR is more accurate with strong sensitivity and specificity for the detection of H. pylori in laryngeal mucosa , but it is less available and of high cost, limiting its use.
H. pylori infection was considered to cause epithelial cell proliferation in the laryngeal mucosa as its effect in gastric mucosa, which eventually could progress to laryngeal carcinoma .
Early treatment of H. pylori is important to decrease the incidence of occurrence of laryngeal pathology especially malignant lesions because it could be considered as a risk factor of cancer larynx according to this study, which reflects significant association between H. pylori and cancer larynx. Accordingly, the treatment of H. pylori should be continued after surgery to help avoid recurrence. Other studies are needed to determine the effect of treatment for H. pylori on recurrence of laryngeal lesions. Moreover, another study is needed to examine this organism within tissue samples of recurrent cases.
| Conclusion|| |
Laryngeal lesions are commonly associated with H. pylori infection, which showed significantly higher level in laryngeal SCC than benign lesions, reflecting more association of larynx cancer with H. pylori infection.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| References|| |
Sheikhian A, Ataherian S, Delfan M, Ebrahimzadeh F, Pournia Y. Prevalence and risk factors of Helicobacter pylori
infection among health center referrals in Khorramabad West of Iran. Asian J Epidemiol 2011; 4:1–8.
Malfertheiner P, Megraud F, O’Morain CA, Atherton J, Axon AT, Bazzoli F et al.
European Helicobacter Study Group. Management of Helicobacter pylori
infection − the Maastricht IV/Florence Consensus Report. Gut 2012; 61:646–664.
Yokoyama A, Yokoyama T, Omon T et al. Helicobacter pylori
, chronic atrophic gastritis.inactive aldehyde dehydrogenase-2, macrocytosis and multiple upper aerodigestive tract cancers and the risk for gastric cancer in alcoholic Japanese men. J Gastroenterol Hepatol 2007; 22:210–217.
Akbayir N, Basak T, Seven H, Sungun A, Erdem L. Investigation of Helicobacter pylori
colonization in laryngeal neoplasia. Eur Arch Otorhinolayngol 2005; 3:170–172.
Tominaga S. Major avoidable risk factors of cancer. Cancer Lett 1999; 143:S19–S23.
Zhuo XL, Wang Y, Zhuo WL, Zhang XY. Possible association of Helicobacter pylori
infection with laryngeal cancer risk: an evidence-based meta-analysis. Arch Med Res 2008; 39:625–628.
Ozyurt M, Gungor A, Ergunay K, Cekin E, Haznedaroglu T. Real-time PCR detection of Helicobacter pylori
and virulence-associated cagA in nasal polyps and laryngeal disorders. Otolaryngol Head Neck Surg 2009; 141:131–135.
Yamamura F, Yoshikawa N, Akita Y, Mitamura K, Miyasaka N. Relationship between Helicobacter pylori
infection and histologic features of gastritis in biopsy specimens in gastroduodenal diseases, including evaluation of diagnosis by polymerase chain reaction assay. J Gastroenterol 1999; 34:461–466.
Fauci AS, Braunwald E, Kasper DI, Hauser SL, Longo DL, Jameson DR, Loscalzo J. editors. Harrison’s principles of internal medicine, 17th edition (Harrison’s principles of internal medicine (single vol.). New York, NY: McGraw-Hill Medical; 2008.
Kusano K, Inokuchi A, Fujimoto K, Miyamoto H, Tokunaga O, Kuratomi Y et al.
Coccoid Helicobacter pylori
exists in the palatine tonsils of patients with IgA nephropathy. J Gastroenterol 2010; 45: 406–412.
Barakat G, Nabiel Y, Ali O, El-Nady G, Musaad A, El-Sharkawy A. Urea and cag A genes of Helicobacter pylori
in Egyptian patients with laryngeal squamous cell carcinoma and benign laryngeal polyps: a cohort study. Eur Arch Otorhinolaryngol 2016; 273:3243–3248.
Siupsinskiene N, Jurgutaviciute V, Katutiene I, Janciauskas D, Vaitkus S, Adamonis K. Helicobacter pylori
infection in laryngeal diseases. Eur Arch Otorhinolaryngol 2013; 270:2283–2288.
De Korwin JD. Advantages and limitations of diagnostic methods for H. pylori
infection. Gastroenterol Clin Biol 2003; 7 (Pt 2):380–390.
Urita Y, Hike K, Torii NL, Kikuchi Y, Kurakata H, Kanda E et al.
Comparison of serum IgA and IgG antibodies for detecting Helicobacter pylori
infection. Intern Med 2004; 43:548–552.
Rezaii J, Tavakoli H, Esfandiari K, Ashegh H, Hasibi M, Ghanei M et al.
Association between Helicobacter pylori
infection and laryngohypopharyngeal carcinoma: a case-control study and review of the literature. Head Neck 2008; 30: 1624–1627.
16Fang TJ, Lee LA, Li HY, Yang C, Huang CG. Helicobacter pylori colonization in the larynges of patients with hoarseness. Laryngoscope 2008; 118:389–393.
Akbayir N, Basak T, Seven H, Sungun A, Erdem L. Investigation of Helicobacter pylori
colonization in laryngeal neoplasia. Eur Arch Otorhinolaryngol 2005; 262:170–172.
Aygenc E, Selcuk A, Celikkanat S, Ozbek C, Ozdem C. The role of Helicobacter pylori
infection in the cause of squamous cell carcinoma of the larynx. Otolaryngol Head Neck Surg 2001; 125:520–521.
Gong H, Shi Y, Zhou L, Tao L, Shi Y, Cao W, Cheng L. Helicobacter pylori
infection of the larynx may be an emerging risk factor for laryngeal squamous cell carcinoma. Clin Transl Oncol 2012; 14:905–910.
Titiz A, Ozcakir O, Ceyhan S, Yilmaz YF, Unal A, Akyon Y. The presence of Helicobacter pylori
in the larynx pathologies. Auris Nasus Larynx 2008; 35:534–538.
21Said RM, Cheah P-L, Chin S-C, Goh K-L. Evaluation of a new biopsy urease test: Pronto Dry, for the diagnosis of Helicobacter pylori
infection. Eur J Gastroenterol Hepatol 2004; 16:195–199.
Nurgalieva ZZ, Graham DY, Dahlstrom KR, Wei Q, Sturgis EM. A pilot study of Helicobacter pylori
infection and risk of laryngopharyngeal cancer. Head Neck 2005; 27:22–27.