Table 2. Summary of Single Cell Analysis... | American Society of Hematology

Table 2.

Summary of Single Cell Analysis of Six Cases of T-Cell–Rich B-Cell Lymphoma

Patient	Experiment	Cells Positive	PCR Products		Rearrangements		Controls Positive
Patient	Experiment	Cells Positive	Total	Sequenced	Repeated	Unique	Controls Positive
1	a	25/32	16VH4L^*	16	16VH4		0/13 buffer
			24Vκ3	23	16Vκ3(a)
					5Vκ3(b)
					2Vκ3(a and b)
2	a	16/33	10VH3L	10	10VH3		0/6 T cells
			14Vκ1	9	8Vκ1(a)		0/13 buffer
					1Vκ1(a and b)
	b	13/30	5VH3L	5	5VH3		0/12 buffer
			13Vκ1	12	9Vκ1(a)
					3Vκ1(b)
3	a	36/60	19VH4L	16	16VH4		0/24 buffer
			33Vκ2	21	21Vκ2
4	a	13/20	10Vκ1	9	9Vκ1		0/8 buffer
			6Vκ2	6	6Vκ2
			0VHL
	b	29/39	23VH3	23	23VH3		0/17 buffer
			28Vλ1	20	20Vλ1
			28Vλ3	6	6Vλ3
5^†	a	34/63	0VHL				0/24 buffer
			10VH4	10	10VH4
			10Vκ4	4	4Vκ4
			20Vλ2	20	20Vλ2
6^‡	a	15/30	4VH1L	4	4VH1		0/12 buffer
			12VH4L	12	11VH4
						1VH4
			1VH3L	1		1VH3
			3Vκ1	3		3Vκ1^2-153
			1Vκ2	1		1Vκ2
	b	18/29	10VH1L
			7VH4L
			11Vλ1	11	11Vλ1		0/14 buffer
			18Vλ3	8	8Vλ3

Patient	Experiment	Cells Positive	PCR Products		Rearrangements		Controls Positive
Patient	Experiment	Cells Positive	Total	Sequenced	Repeated	Unique	Controls Positive
1	a	25/32	16VH4L^*	16	16VH4		0/13 buffer
			24Vκ3	23	16Vκ3(a)
					5Vκ3(b)
					2Vκ3(a and b)
2	a	16/33	10VH3L	10	10VH3		0/6 T cells
			14Vκ1	9	8Vκ1(a)		0/13 buffer
					1Vκ1(a and b)
	b	13/30	5VH3L	5	5VH3		0/12 buffer
			13Vκ1	12	9Vκ1(a)
					3Vκ1(b)
3	a	36/60	19VH4L	16	16VH4		0/24 buffer
			33Vκ2	21	21Vκ2
4	a	13/20	10Vκ1	9	9Vκ1		0/8 buffer
			6Vκ2	6	6Vκ2
			0VHL
	b	29/39	23VH3	23	23VH3		0/17 buffer
			28Vλ1	20	20Vλ1
			28Vλ3	6	6Vλ3
5^†	a	34/63	0VHL				0/24 buffer
			10VH4	10	10VH4
			10Vκ4	4	4Vκ4
			20Vλ2	20	20Vλ2
6^‡	a	15/30	4VH1L	4	4VH1		0/12 buffer
			12VH4L	12	11VH4
						1VH4
			1VH3L	1		1VH3
			3Vκ1	3		3Vκ1^2-153
			1Vκ2	1		1Vκ2
	b	18/29	10VH1L
			7VH4L
			11Vλ1	11	11Vλ1		0/14 buffer
			18Vλ3	8	8Vλ3

The micromanipulated cells were analyzed together with buffer or T-cell controls in a blinded fashion. The data summarized as one experiment contain all data obtained from cells and controls micromanipulated in one micromanipulation session. Two different V_κ rearrangements using V_κ genes of the same V gene family were amplified from cells of patients 1 and 2. The different rearrangements are labeled with the suffixes (a) and (b) in Tables 2 and 3.

Ig heavy chain rearrangements with the suffix ‘L’ were amplified with the V_H leader primers.

†

For 20 cells the V_HL and V_κ primer sets were used, for 43 cells the V_HFRI and the V_λset.

‡

In experiment b of case 6 the V_HL and V_λprimer sets were used, but nested PCR for V_HL was only performed for V_λ positive cells.

F2-153

An unmutated V_κL12a in-frame rearrangement was coamplified with the clonal V_H4 rearrangement from cell BKL1. As no V_κ rearrangements were amplified from the 10 other cells from which clonal V_H rearrangements were obtained, the amplification of this unique V_κ1 rearrangement is most likely due to contamination with fragments from other cells during the micromanipulation procedure.

View Large

This Feature Is Available To Subscribers Only