FİTOHEMAGGLÜTİNİN KONSANTRASYONUNUN DOWN SENDROMLU VE SAĞLIKLI BİREYLERİN LENFOSİTLERİNDEKİ MİTOTİK İNDEKSLERİNE ETKİSİ

İnsan periferal kan T-lenfositlerinin mitoza yönlendirilmesi ve kromozom analizlerinde birim hacimdeki büyüme ortamına eklenen fitohemaglütinin (FHE) miktarı bir laboratuardan diğerine göre önemli ölçüde değişmektedir. Bunun nedeni bilinmemekte ve literatürde de bu konuda yeterli bir bilgiye ulaşılamamaktadır. Alışılmış periferal kan kültürü (72 saat) ve kromozom hazırlama işlemi, yaşları 0-8 yıl arasında değişen Down Sendrom (DS)’lu hasta (N=30) ve sağlıklı kontrollerinden (N=24) alınan kan örneklerinin, aynı fakat konsantrasyonu gittikçe artan şekilde: 0.37 ml, 0.75 ml, 1.48 ml ve 2.21 ml FHE /100 ml’lik ortam’lara ekilmesi ile yapılmışlardır. FHE’nin dört farklı konsantrasyonunu içeren dört farklı ortamdan elde edilen her bir preparatta Mitotik indeks (Mİ)’ler, birey ve konsantrasyon başına 5000- 6000 uyarılmış lenfositler sayılarak yüzde (%) olarak belirlendi. Hem DS’lu, hem de kontrol gruplarının genel ortalamaları alındı. Bireyler arasında önemli farklılıklar görülmesine karşın, kültür ortamında FHE konsantrasyonunun artış yönüne doğru (%0.37, %0.75, %1.48, ve %2.21) lenfositlerdeki Mİ’lerin genel ortalamaları karşılıklı olarak; 30 DSlu’da %4.20±1.73, %4.57±1.31, %4.69±1.19 ve %4.19±1.11 iken 24 kontrolde %4.37±1.67, %5.06±1.60, %5.01±1.47 ve %4.98±1.43 olarak bulundu. Sonuçta, Biological İndustries (İsrail) ürünleri kullanıldığında, % 1-1.5 hacim/hacim FHE, hem DS’lu, hem de kontrollerin kan kültürleri için optimum (hem etkin hem de tutumlu) bulundu

Anahtar Kelimeler:

Down sendromu, fitohemaglütinin (FHE), FHE konsantrasyonu, lenfosit, mitotik indeks

Effect of the Phytohemagglutinin Concentration on the Mitotik Indexes of Down Syndrome and Healthy Individuals’ Lymphocytes

The amount of phytohemagglutinin (PHA) amount added to the growing medium in unit volume of human peripheral blood T-lymphocytes induced mitosis and at chromosome analysis varies significantly from one laboratory to another. The reason for this variation is not known, and in literature adequate information on this subject is not available. Conventional peripheral blood culture (72h) and chromosome preparation procedure have been used except that blood samples from Down syndrome (DS) patients (N=30) and healthy controls (N=24), with the age range of 0-8 years, have been cultivated in the same but gradiently increasing PHA concentration containing: 0.37 ml, 0.75 ml, 1.48 ml and 2.21 ml of PHA / 100 ml of medium. Mitotic indexes (MIs) of each specimen obtained from four separate media with four different concentrations of PHA were determined as percentage (%) by screening from 5.000 to 6.000 stimulated lymphocytes per concentration per individual. Overall means MIs of both DS and control groups were taken. Although considerable interindividual variations were apparent, overall means of MIs in lymphocytes according to the gradient of PHA concentration (0.37%, 0.75%, 1.48%, and 2.21%) in culture medium were found as 4.20±1.73%, 4.57±1.31%, 4.69±1.19%, and 4.19±1.11% for 30 DS patients and 4.37±1.67%, 5.06±1.60%, 5.01±1.47%, and 4.98±1.43% for 24 healthy individuals, respectively. In conclusion, When the Biological Industries (Israel) products are used, 1-1.5 % volume/volume PHA was found optimum (both effective and economic) for the blood cultures of both DS patients and healthy controls

Keywords:

Down syndrome, lymphocyte, mitotic index, phytohemagglutinin (PHA), PHA concentration,

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