Bes1 Dna In Bd Skin Lesions

The 999 bp fragment of Bes-1 DNA was detected by PCR in mucocutaneous lesions from patients with BD (Fig. 2). The product yielded the expected fragments when digested with Pst-1 restriction endonuclease (Fig. 3). Among the BD patients, 37% were positive for Bes-1 DNA (three of 11 cases) and this was confirmed using skin samples without epidermis (Fig. 3, Table 2). We also demonstrated by PCR the presence of Bes-1 DNA in samples of a genital ulcer (No. 4, incomplete type) and an EN-like eruption (No. 22, complete type) from patients with BD, and Bes-1 DNA was found in the skin sample of an EN-like eruption (No. 7) and an oral aphtha (No. 8) in one patient with BD (suspected type). Bes-1 DNA was also detected by PCR-ISH in the muco-cutaneous lesions, which were positive by PCR (Table 2). The reaction product was deposited on the nuclei of cells adhering to the dermal vessel walls and some of the infiltrates around the vessels in the main inflammatory lesions (Fig. 4). The PCR-ISH-positive cells correlated with the presence of Bes-1 DNA in the lesions from patients with BD.

Table 1. Details of patients with Behget's disease (BD) and other inflammatory disorders

No./Sex/Age

Diagnosis

Location

Duration and symptoms l/F/44y 2/F/30y 3/M/29y* 4/M/29y* 5/F/48y 6/F/23y 7/F/47y** S/F/47y** 9/F/23y 10/M/56y U/F/70y 12/F/29y 13/M/49y 14/M/50y*** 15/M/50y*** 16/M/50y*** 17/F/41y 18/F/23y 19/F/28y 20/F/25y 21/F/39y 22/F/49y

Sweet's disease lower leg 2mo: myelodysplastic syndrome

BD lower leg 2y: EN-like symptom

BD scrotum 5y: genital ulcer

BD scrotum 5y: genital ulcer

EN lower leg 2w:

EN lower leg lw:

BD lower leg Imo: EN-like symptom

BD oral mucosa lmo: oral aphthae

Phlegmone lower leg 2w:

Sweet's disease chest 3w:

EN lower leg lmo: drug-induced

BD arm 7y: EN-like symptom

BD scrotum 4y: genital ulcer

BD trunk 3y: folliculitis

BD finger 3y: folliculitis

BD finger 3y: folliculitis

Sweet's disease lower leg 2.5mo: hyperthyroidism

BD lower leg 2.5y: EN-like symptom

BD upper leg 1.5y: EN-like symptom

BD Lip 7y: oral aphthae

BD lower leg 4y: EN-like symptom

BD arm 24y: EN-like symptom

*,**,***; three patients have several lesions, EN: erythema nodosum

Figure 3. Bes-1 DNA in mucocutaneous lesions. A) The epidermis was removed from Bes-1 positive samples by micro-dissection. Representative data are shown (No.4 and 7). Lanes 4 and 7 include epidermis and lanes 4' and 7' are without epidermis. Positive bands were detected in all lanes. B) Electrophoresis of restriction endonuclease digested DNA. Representative data are shown (No.7). Positive control, Bes-1 and No.7 showed the expected fragment pattern; uncut 999-bp sample and sample cut with Pst-1 showing 160- 389- and 450-bp fragments.

Figure 3. Bes-1 DNA in mucocutaneous lesions. A) The epidermis was removed from Bes-1 positive samples by micro-dissection. Representative data are shown (No.4 and 7). Lanes 4 and 7 include epidermis and lanes 4' and 7' are without epidermis. Positive bands were detected in all lanes. B) Electrophoresis of restriction endonuclease digested DNA. Representative data are shown (No.7). Positive control, Bes-1 and No.7 showed the expected fragment pattern; uncut 999-bp sample and sample cut with Pst-1 showing 160- 389- and 450-bp fragments.

Figure 4. Bes-1 DNA in muco-cutaneous lesion. PCR-1SH analysis of EN-like eruption in one BD case (No. 7) shows the positive signals (arrows) for Bes-1 DNA in cells adhering to the superficial dermal vessel wall (left), and deep dermal vessel walls (right).

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