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Table 1: Xenopus Lymphocyte surface markers detectable with available mAbs
| Xenopus markers (mAbs) | Expression Pattern |
|---|---|
| CD8 (AM22, F17) |
Larval and adult thymocytes (70-80%) and T-cells (about 20% of splenocytes). All lymphoid tumor lines |
| CTX (X71, 1S9.2) |
Larval and adult thymocytes (60-70%)no consistent expression in periphery. All lymphoid tumor lines and gut epithelial tissue. |
| XT1 (XT1) |
Most, but not all, larval and adult T-cells; earliest marker of thymocytes. All lymphoid tumor lines |
| MHC class I (TB17) |
Ubiquitous in adult; all lymphopoietic lineages. Not expressed until metamorphosis |
| MHC class II (AM20, 14A2) |
Thymocytes, B and T-cells (99% of spleen lymphocytes), only B-cells in larvae |
| CD5 (2B1) |
Thymocytes (>95%), T-cells and some PMA-activated IgM+ B cells. All lymphoid tumor lines |
| NK-like (1F8) |
Non-B and non-T, peripheral lymphoid cells. |
| RC47 | Leukocyte lineage from very early stage. Thymic cortex and medulla (>90% of total thymocytes) |
| IgM (10A9) |
Larval and adult B cells. |
| IgY (11D5) |
Some larval and adult B cells. |
| IgX (410D9) |
Some larval and adult B cells, especially in the gut. |
Table 2: Some available cDNA encoding immunologically-relevant X. laevis genes
| Immune System Characteristics | Larva (Ancestral-like system) Adult (Mammalian-like system) |
|---|---|
| CTX | Thymus, all lymphoid tumor lines and gut epithelial tissue. |
| TCR α, β, γ | thymus, circulating T-cells |
| BCR μ, γ, ξ, L | B cells |
| MHC class I | ubiquitous in adult |
| Non-classical class Ib | more restricted tissue distribution for some of them MHC class II |
| MHC class II | thymus, splenic lymphocytes |
| Rag 1 & 2 | mainly thymus, at lower level in bone marrow, kidney, spleen and liver |
| TdT | mainly thymus of old larvae and adult, less in spleen |
| Ikaros | mainly thymus |
| GATA III | thymus and T-cells |
| LMP7 | Thymus, spleen |
| Hsps (gp96, hsp70) | Ubiquitousely expressed, |
Table 3: Currently available X. laevis MHC-defined stains and clones
| Name, (MHC gentotype) | |
|---|---|
| Partially inbred, MHC homozygous strains | F, J |
| Isogenetic leavis/gilli (LG) clones with identical heterozygous (a/c) MHC genotype | LG-6, LG-7, LG-15 |
| MHC-disparate LG isogenetic clones | LG-3 (b/d), LG-5 (b/c) |
Table 4: Available cloned thymic tumor cell lines
| Name of Tumor Cell Line | BB7 | ff-2 | 15/0 |
|---|---|---|---|
| Genetic background of initial tumor-bearing host | Partially inbred MHC homozygous ff strain |
LG-15 (MHC a/c) isogenetic clone | |
| T-cell surface markers (CD8, CD5, pan T-cell) | +++ | +++ | +++ |
| Ig mRNA | - | + | + |
| Ig protein | - | - | - |
| MHC class I mRNA and protein | - | + | - |
| MHC class II mRNA and protein | - | - | - |
| Tumorigenicity in syngeneic | |||
| Larvae | - | + | ++ |
| Adult | |||
List of available genomic and cDNA libraries
| X. laevis cDNA libraries |
|
|---|---|
| Old libraries in λZAPII vector | |
| ST: | Outbred adult spleen + thymus (constructed by L. Du Pasquier, Basel Switzerland)) |
| LG7Ad: | Adult from the clone LG7 immunized with DNP-KLH + CFA (constructed L. Du Pasquier) |
| LG7 Tadp: | LG7 pre-metamorphic larvae immunized DNP-KLH + alum (constructed by M. Wilson, Jakson MS) |
| SMART cDNA libraries (λZAP-express, constructed by S. Guselnikov and A. Taranin, Novosibirsk, Russia) |
|
| T: | Outbred larval spleen + thymus + liver. 7x105 independent clones, titer 6x109 |
| S: | Outbred adult spleen. 2x106 independent clones, titer 1x109 |
| NK: | NK cells, 1F8+ purified from outbred adult splenocytes. 3x106 independent clones, titer 1x1010 |
| T: | CD8 NK/T cells (AM22+/1F8+) purified from outbred splenocytes. 5x105 independent clones, titer 4x109 |
| Arrayed EST cDNA libraries (constructed by Express Genomics, Inc) |
|
| Outbred adult thymus | |
| Outbred adult splenocytes activated for 4 days with PHA | |
| X. tropicalis cDNA libraries (λZAP-express) |
|
| Trop: | Adults spleen + thymus + intestine. Titer 5x1010 |
Table 5: Usual Primer Pairs for RT-PCR
| Activation-induced cytosine-deaminase (AID); | |
| Forward: | 5’-TCA CGA CCC CCA TAG GAA CTA C-3’ |
| Reverse: | 5’-GGA TTT AGG AGA CTT TGC CTC AAG-3’ |
| TM: | 58ºC; product length: 450 bp (span intron) |
| β-actin: | |
| Forward: | 5’-GGT GTC ATG GTT GGA ATG G-3’ |
| Reverse: | 5’-TGG GTT ACA CCA TCA CCT GAG-3’ |
| TM: | 56ºC; product length: 359 bp |
| β2-microglobulin | |
| Forward: | 5’-CCG GTG GTC AAG GTT TAC ACT G-3’ |
Reverse: |
5’-AGC TGT GAA AGA TTA AAC CCT-3’ |
| TM: | 57ºC; product length: 800, 950 bp (2 bands correspond to the 2 X. laevis genes; primers also span an intron) |
| α2-macroglobulin receptor CD91: | |
| Forward: | 5’-AAT GGA GAC ATC ATC TGC TCG TG-3’ |
| Reverse: | 5’- CAG GGT CGC ATA GAC TGG ATT G-3’ |
| TM: | 56ºC; product length: 500 bp (span intron) |
| Chytrid Fungus ribosomal RNA Intervening Transcribed Sequence (Batrachochytrium dendrobatidis ITS) |
|
| Forward: | 5’- CCT TGA TAT AAT ACA GTG TGC CAT ATG TC-3’ |
| Reverse: | 5’- AGC CAA GAG ATC CGT TGT CAA A-3’ |
| TM: | 60ºC; product length: 250 bp (span an intron) |
| CTX: | |
| Forward: | 5’-GCA GCA GCG GTA ATC GGA G-3’ (position 214) |
| Reverse: | 5’-CTC AGC ATG GTC ATG GAA TTG-3’ (position 287) |
| TM: | 56ºC; product length: 225 bp (1.3 kb if genomic DNA amplified) |
| Elongation factor α1 (Ef-α1): | |
| Forward: | 5’-CCT GAA TCA CCC AGG CCA GAT TGG TG-3’ |
| Reverse: | 5’-GAG GGT AGT CTG AGA AGC TCT CCA CG-3’ |
| TM: | 56ºC; product length 223 bp |
| Frog virus 3 Immediate early protein (IE-FV3): | |
| Forward: | 5’- ATG ATC CAA GCC TAC CTG TGC-3’ |
| Reverse: | 5’- AAA TGT CCT AAT CTA TAC ACC-3’ |
| TM: | 51.5ºC; product length 420 bp |
| Frog virus 3 major capsid protein (MCP): | |
| Forward: | 5’- GAC TTG GCC ACT TAT GAC -3’ |
| Reverse: | 5’- GTC TCT GGA GAA GAA GAA-3’ |
| TM: | 55ºC; product length 520 bp. Remark: primers work also with ATV |
| Gp96: | |
| Forward: | 5’- GTTGTAAGCAGGGAAGAGG-3’ |
| Reverse: | 5’-TGTCTCAGTCTTGCTGCTCCACAC-3’ |
| TM: | 56ºC; product length 500 bp |
| HSC-70 | |
| Forward: | 5’-TGG AGT AGA AAG GAA TGT ACT GAT C-3’ |
| Reverse: | 5’-AAC TTC ATT GCA TTT CTC CAG-3’ |
| TM: | 58ºC; product length: 747 bp |
| HSP70 | |
| Forward: | 5’-AGA CTG ATC GGG AGA AAG TTC-3’ |
| Reverse: | 5’-CTT TTC CAC AGG TTC CAA GGT ACC C-3’ |
| TM: | 58ºC; product length: 747 bp |
| Immunoglobulin heavy chain μ, constant domain (IgH-Cμ): | |
| Forward: | 5’- GCA ATG CCA AAC ACC TGG-3’ |
| Reverse: | 5’- TCT TCT CTT GTA AGG AAC CCG-3’ |
| TM: | 58ºC; product length 350 bp |
| Immunoglobulin heavy chain γ, constant domain (IgH-Cγ): | |
| Forward: | 5’- ATC CAG CAG CCA ACT CAC CAT C -3’ |
| Reverse: | 5’- AAG GTG GCA GCA AGG TGA TTG -3’ |
| TM: | 58ºC; product length 720 bp |
| Interleukin 1β (IL-1β): | |
| Forward: | 5’- AAC AGA AGA TGG CCA AGA CTC-3’ |
| Reverse: | 5’- ATG CAA CCG ATT CAA AGC TG-3’ |
| TM: | 55ºC; product length:152 bp. (span an intron) |
| Low mass protein 7 (LMP7): | |
| Forward: | 5’- CAG AAT TCG CNG AYT CYC ART AYT GG -3’ |
| Reverse: | 5’- CAG AAT TCC CAN CCR TCY TCY TTC AT-3’ |
| TM: | 53ºC; product length 400 bp |
| Major histocompatibility antigen class I (MHC class I): | |
| Forward: | 5’-GAGTTCTTTGCCCTGGATACA-3’ (Lineage A α2-specific) |
| Forward: | 5’-GAGTTCATAGCGCTGGATACA-3’(Lineage B α2-specific) |
| Reverse | 5’-CACAATAAGTTTCTCCTTCAGG-3’(Lineage A, and B specific) |
| TM: | 59ºC; product length: 450 bp |
| Major histocompatibility antigen class IIα (MHC class IIα): | |
| Forward: | 5’-TCAAAGTCAGTGGTTTGGACG -3’ |
| Reverse: | 5’-GTGTAAATATCATGTTCATTTGGT-3’ |
| TM: | 58ºC; product length: 390 bp |
| Major histocompatibility antigen class class IIΒ (MHC class IIΒ): | |
| Forward: | 5’-ACGGCACCGACAATGTCAGG -3’ |
| Reverse: | 5’-TTAATGGTTGTCTCCAGCATC-3’ |
| TM: | 58ºC; product length: 450 bp |
| Recombination activating gene 1 (Rag 1): | |
| Forward: | 5’- GCG CCA AGA ATCTGT GTC ACT-3’ |
| Reverse: | 5’- GAT AGC TCC GGT TCT GCT GAT-3’ |
| TM: | 58ºC; product length: 446 bp |
| Recombination activating gene 2 (Rag 2): | |
| Forward: | 5’- GAG CAC AGA GGG AAG AAC AG-3’ |
| Reverse: | 5’- (GCA CAC AGC TGG ATG CCT AA-3’ |
| TM: | 56ºC; product length: 298 bp |
| Terminal deoxynucleotidal transferase (TdT): | |
| Forward: | 5’- ATC CAC TGA GCC AAT-3’ |
| Reverse: | 5’- CCT TCA TCG CTG TTA-3’ |
| TM: | 56ºC; product length: 616 bp |
Ordering Info
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Reource Quick-Links:
Surface Markers
cDNA encoding
Stains & Clones
Cloned thymic tumor cell lines
genomic and cDNA libraries
Primer Pairs for RT-PCR
