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1. Tarlo et al 1993 Weber 1993 Wuhrich 1993 Nish et al 1991
2. Tarlo etal 1993 Weber 1993 Wuhrich 1993 Nish er al 1991
3. 10 10 nM WEHI 279 10 FBS RPMI 1640 72 Table 4 7 10 nM 100 nM Table 4 7 Dose dependent Effect of Eugenol on Immunoglobulin Production by WEHI 279 Cells sample Concentration Relative Ig content nM IgM IgG Control 0 1 0 1 0 Eugenol 10 1 0 0 9 10 0 9 i 10 0 9 t i 109 1 0 1 2 10 0 9 1 1 10 0 1 0 2 10 0 0 Ig concentrations of the control culture were 24 7 ug ml for Ig M and 11 3 ng ml for IgG 4
4. 1993 Westerholm et al 1995 Zapponi et al 1997 DEP lgE Diaz Sanchez et al 3 1994 Takenaka et al 1995 PAH Holladay et al 1994 2 3 7 8 Tetrachlorodibenzo p dioxin TCDD B Karras et al 1996 Rin LORETO FVM TST DERE lgE Metcalfe 1991 1gA lgsG lgE
5. Fig 3 1 10 10 uM IgG IgM Wis Sei fae regen ME dC Y UST ciae FCFIX10 2 510 nM IlgE 10 nM IgE IgE IgG IgM Amaranth New Coccine Tartrazine Rela v E Concentration Sunset Yellow FCF Food Coloring uM Fig 3 1 Effect of Azo Colorings on Immunoglobulin Production by Rat Spleen Lymphocytes Spleen lymphocytes were cultured with various azo dyes at around 10 to 10 uM for 72 hr and the culture supernatant was collected to evaluate Ig concentration Relative Ig Concentration Food Coloring uM Fig 3 2 Effect of Azo Colorings on Immunoglobulin Production by Rat MLN Lymphocytes MLN lymphocytes were cultured with various azo dyes at around 10 to 10 uM for 72 hr and the culture supernatant was collected to evaluate g concentration MLN
6. IgE Helbling et al 1993 33 i EE Niinimaki et al 1995 IgE Metcalfe 1991 lgA IgG IgE 1
7. 0 4 49 units Taqd Fermentas Vilnius Lithuania 2 5 mM 200 LM 10 uM cDNA Gene Amp PCR system 2400 Perkin Elmer Foster City CA PCR 2 Sawady Technology DNA Kodak Digital Science Eastman Kodak Rochester NY SOIR Duncan Duncan 1955 wT ICs SSH 1 3y RERI UDANKOMABEICRIZT FUTT JERONE
8. Curcuma 7ojga Li er al 1993 Sui etal 1993 Yasuni et al 1993 Soni et al 1992 Fujiyama Fujiwara et al 1992 Ammon et al 1993 Srivastava et al 1995 Huang et al 1991 Kakar et al 1994 Lu et al 1994 TPA Yasukawa et al 1994 17 ine Oey VV Qv 858 TAT VV eS LC LA eee WERVENDE TORINIZCKBW CULT UN TY HRNILEORB SE Metcalfe 1991 IgA
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10. DEP Muranaka et al 1986 Takafuji er al 1987 1989 Takenaka et al 1995 B Savage et al 1991 Fischer etal 1994 BB
11. 10 MLN lsG IgM ORAS TR AB EOS ARF Vy CD FCF 10 10 uM IgE IgE IgG IgM Water soluble Annatto Brilliant Blue FCF Cu chlorophyllin Fast Green FCF Relative lg Concentration Indigocarmine Food Coloring uM Fig 3 3 Effect of Non Azo Colorings on Immunoglobulin Production by Rat Spleen Lymphocytes Spleen lymphocytes were cultured with various colorings such as triphenylmethane dyes brilliant blue FCF and fast green FCF indigoid dye indigocarmine water soluble annatto and sodium Cu chlorophyllin at around 10 to 10 uM for 72 hr and the culture supernatant was collected to a evaluate Ig concentration IgE IgG IgM g Concentration Relati e V Food Coloring uM Fig 3 4 Effect of Non Azo Colorings on Immunoglobulin Production by Rat MLN Lymphocytes MLN lymphocytes were cultured with various colo
12. Schwartzer et al 1992 Cu Albertini et al 1996 Fe Cu IgE 4 1 Tarlo et al 1993 Weber
13. 1 mM IgG IgM lgG IgM 10 uM 100 uM 20 Table 3 1 Dose dependent Effect of Water soluble Colorings on Immunoglobulin Production by Rat Spleen Lymphocytes Coloring Control Cacao pigment Cochineal pigment Corn pigment Betanin Carthamus yellow Monascus pigment Concentration uM 10 100 1000 10 100 1000 10 100 1000 10 100 1000 10 100 1000 10 100 1000 Relative Ig content IgE 1 00 0 07 0 74 0 0 7 0 88 0 08 0 71 0 04 0 70 4 07 0 78 0 11 0 80 0 04 Q 7220 04 0 78 0 07 1 09 0 16 0 43 0 02 0 5440 05 0 7620 09 0 69 0 09 0 89 0 05 0 83 0 04 0 58 0 0 3 0 81 0 06 1 26 0 04 IgG 1 00 0 03 9620 02 1 03 0 02 02 0 01 0 99 0 04 1 08 0 03 7080 04 Q 95 0 03 1 03 0 04 Dg ON OT S 0 94 0 02 0 91 0 04 GZ 7002 0 92 0 01 1 02 0 04 Q0a0 00 0 97 0 02 0 9Z 0 03 0 09 0 06 IgM 1 0020 03 0120520102 I DL ego T I lad b 3 1 06 0 02 ZIOZP 0 929 ZEN 1 02
14. Table 4 1 IgM 10 ng ml Table 4 1 Dose dependent Effect of Ethanol Extracts of Spices on IgM Production by Mouse Spleen Lymphocytes Relative IgM content Spice 107 fo WO 10 iO pemi Allspice powder 1 0 0 89 POTS POR VWI 1 95 Onion powder OQ L03 Q91 0 91 Q97 0 86 Garlic 1 07 085 Oreo 0 85 01079 665 Curry 1 4 1 1 1 0 Loy 10 0 81 Clove powder 12 0 91 0 82 0 88 0 89 1 05 Pepper black Teed 0 75 0 88 0 89 O86 0 88 Pepper white 1 2 092 Loy DOs 45d 0 95 Horseradish 1 1 091 OO 0 95 0 29 0 91 Cinnamon 157 POON TOS L092 097 G15 Ginger 0 94 0 88 0 90 0 79 0 68 0 32 Sage powder 1 2 0 87 0 90 O97 O75 0 08 Thyme powder 1 08 1202 Oe O34 WYSE c9 Chili powder i 093 0 96 096 Gee 13 Red pepper 1 1 4 LOL lee 0 860 0 97 Nutmeg lll Ose 0 86 LOI 085 0 47 Paprika powder 0 96 1 6 0 95 1 4 py 1 05 107 IgM concentration of the control culture was 1 3 mg ml 36 OU gue micis v Cte API AIT AAR A HM VIS PAYED ASO REDD La APUA
15. Warner et al 1979 Gutman er al 1981 IlgG IgE Fig 2 3 B autocrine paracrine B Pistoia 1997 WEHI 279 IgM IgG IgE Kindler et al 1997
16. 1gG IgE 1 AMF CULT Y bH EK OMe DY OSE VY NERO BAO FS EE IT RIE eb 2 Bh 1 6 Yo Ay EE ars MESE wm dS K UEB PARAS 8 MOIR Metin VER Sete SF ee Se Sean OD AMR ju ue OI Ra BOA 9158 9A Pee ACR ep rm SI e SP o PPSS KGET ISAM ETS FOF Sam AV DABIT UV Sp FCF PBS DH 7 4 1710 DMSO 1 1000 LE 0 05 Tween 20 PBS TPBS
17. Lim et al 1994a MLN RPMI 1640 mid cds X OMLNz T V DELTY Lympholyte Rat Cedarlane Hornby Canada 10 Intergen Purchase NY RPMI 1640 2 x 10 cells ml lIgE 6 IgG IgM 72 10 CO 3 Yamada etal 1993 IgG lgM IgE 96 37C 1 TPBS 3
18. IgE 4 Y 52 VADER PE POO pie Hy OS BL do Rs EST Lic lgo IgM O COOH Di SEADA Fluorescein Diiodofluorescein Br Br r ON SAN 2 Br Br O ONa QU JJ Eosin B Eosin Y HeC hN O l C4H 4 H5C5 pli ENT yt f B Br SO O 3 Cl COONa C Cl 2 SO4Na CI Acid Red Phloxine I I e I I Wow CI COONa Cl CI CI Erythrosine Hose Bengal Fig 5 3 Structure of Xanthene Dyes 53 2 WEHI 279 1 IgE IgG IgM
19. MLN 10 FBS RPMI 1640 IgE 4 6 IgG IgM 24 72 Lim etal 1994a 1994b CC 7 KRY LOSER RE SAX OX EUM FNSWSNNKm AAV AW TEP CE IgE 6 rgG IgM 72 Fig 5 1 50 uM IgE 30 nM lg IgM 50 4 f IgE 10 mM IgG IgM
20. de WI WE JT AF eats eee re 70 100 ug ml IgG Table 4 2 A SA WV eee er Os Jey Pe 10 ug ml 0 001 ug ml FAT SUUS 0 ea Pa 5239 IgA Table 4 3 A 1 pg ml
21. Yamada et 4L 1996 6 45W of CH CH COOH NH L tyrosine Chg CH COOH NH 3 lodo L tyrosine HO mes xor st an CHy CH COOH NH 2 NH 3 5 Diiodo L tyrosine 3 5 3 Triiodo L thyronine I CHy CH COOH NH Tyroxine b e E lt o e Y Fig 5 9 Structure of Tyrosine Derivatives and Thyroid Hormone B 60 e 10 pM 3 mM pou Y s BL m I uU LS ead EI sd eps lOTuXe rectbkBEEULxosEKUWSHEL DURE E RIS TBE E RE T B
22. 10 ntergen Purchase NY RPMI 1640 2 x 10 cells ml IgE 6 IgG lgM 72 10 CO Tl d al mo iii eee TO B WEHI 279 WEHI 279 10 FBS GIBCO BRL Grand Island NY RPMI 1640 5 CO 1 x 10 cells ml 3 Yamada et al 1993 IgG IgM IgE 96 37 1 TPBS 3
23. IgG IgE B IgM IgG IgE 15 ty Gy Gs chiki et al 1993 Li et al 1994 WEHI 279 Gy Ge IgM IlgG lgE Ahmed et al 1997 Kuramoto et al 1996 Verdaguer et al 1993 WEHI 279 NEd vS OUR T A 3
24. 10 FBS GIBCO BRL Grand Island NY eRPMI 1640 5 CO WEHI 279 1x10 cells ml 10 FBS RPMI 1640 1 2 3 4 6 8 4 WEHI 279 10 FBS PBS FITC IgE IgG IgM 30 10 FBS PBS EPICS PROFILE II Coulter Miami FL 3 RT_ PCR WEHI 279 RNA
25. Table 4 6 100 uM S0 Table 4 0 Dose dependent Effect of Eugenol on Immunoglobulin Production by Mouse Spleen Lymphocytes sample Control Eugenol Ig concentrations of the control culture were 7 5 ug ml for IgM 164 5 ng ml for IgG and 91 7 ng ml for IgA Concentration uM 0 10 10 10 104 10 10 10 107 10 Relative Ig content IgM 1 0 ET 1 1 1 0 1 2 1 142 1 3 0 8 0 IgG 1 0 1 1 1 4 1 0 13 1 1 1 2 his de 0 IgA 1 0 1 0 1 0 1 0 1 0 1 0 1 1 1 1 0 8 0 4 WEHI 279 T B 42 m B
26. 3 AIT Adegoke et al 1994 fr V ROSE Hz RAS REST vci z CRA EB BS 0 8152372 S UC Zheng er al 1992 Rao et al 1995 RIHI k U Capsicum annum Higashimoto et al 1993 Azizan et al 1995 Sastre et al 1996 Cooper et al 1996 Kanerva etal 1996 aniseed Fraj er al 1996 TIA Teu
27. Table 4 4 Dose dependent Effect of Ethanol Extracts of Spices on IgM Production by WEHI 279 Cells Relative IgM content Ba M Spice os FOP qoin o aqu CSS Allspice powder 3 3 1 2 1 5 1 2 1 2 Onion powder 1 3 1 2 Ne 1 2 1 2 1 1 Garlic L2 1 2 1 1 2 1 2 Dy Curry 12 1 2 ii I5 1 06 1 09 Clove powder pho tt 5h uw a o Pepper black E 1 1 Lo7 ll 1 03 1 05 Pepper white 1 2 108 1 1 1 03 21 96 Horseradish 1 07 2408 14 L96 X95 MO Cinnamon 1 1 OG Joy Lay o TO Ginger 1 04 1 1 1 02 1 04 0 98 0 95 Sage powder koed LoS LO LOS X9 1 10 Thyme powder 107 14099 L402 4405 LMO 7086 Chili powder OL d us WD Gre Api Red pepper LOL 109 107 1905 TO 1 0 Nutmeg 1 00 1 1 LOS 108 MQ 0 92 1 02 3404 DOB 31 06 40 OA Paprika powder IgM concentration of the control culture was 5 4 mg ml 40 lgG Table 4 5 0 001 ng ml WEHI 279
28. sO uM 200 20 IgE IgE ng ml IgG ng ml 90 5 c hr d D 10 0 p U E g 2 1 10 0 3 10 10 10 10 40 10 10 10 Food Additive M Fig 5 1 Dose dependent Effect of Food Additives on Immunoglobulin Production by Rat Spleen Lymphocytes Lymphocytes were cultured in the presence of various concentrations of ethyl acetate 6 sodium nitrite O tartrazine Bl Bordeaux S O or Rose Bengal A for 6 h to measure the IgE content and for 72 h to measure the IgG and IgM contents 1 cr 74 y 51 6 72 90 IgE A415 IgG ng ml IgM ng ml Fig 5 2 Effect of Xanthene Dyes on Immuno
29. Lietar 1993 HO lt O CH COOH Caffeic Acid CHO OCH H CH CHCOCH COCH CH OH Curcumin Fig 3 6 Structure of Curcumin and Caffeic Acid Escribano et al 1996 Thresiamma et al 1996 Ursini et al 1994 IgE MLN IgE IL 4
30. 37C 1 4 k TCAE ERE CBI Toe RIT RE UTNE 37C 1 IgE 37C 1 37C 1 37 C 15 415nm 4 RT PCR WEHI 279 RNA 10 PBS TRIzol GIBCO BRL RNA RNA 75 10 ug RNA United States Biochemical Cleveland OH RNA DNA cDNA
31. Table 4 5 Dose dependent Effect of Ethanol Extracts of Spices on IgG Production by WEHI 279 Cells Relative IgG content Spice i19 10 30 309 10 10 gm Allspice powder 1 3 1 2 0 97 1 1 Onion powder 1 3 132 T2 ET 1 08 1 01 Garlic 1 25 1 2 1 1 LI E71 02 Curry 2 2 1 1 1 1 LOSST 10I Clove powder E 109 1 08 104 1 0 Pepper black 1 2 1 1 1 00 1 05 0 97 0 94 Pepper white lez Loo v 02 LOS 10 0 81 Horseradish 12 LA 1 05 105 LOL 104 Cinnamon 1 4 1 4 1 4 1 4 LS 1 3 Ginger 1 4 1 4 i 1 9 1 5 1 2 Sage powder Leo 1 4 1 3 1 3 1 2 1 0 Thyme powder 1 4 1 3 L3 1 2 0 97 MOTE Chili powder 1 4 1 2 i2 LI 1 2 Red pepper Me Lt L3 1 02 1 04 1 08 Nutmeg 12 WZ 1 0 1 0 1 04 1 0 Paprika powder 1 02 1 04 0 98 1 06 1 0 0 94 IgG concentration of the control culture was 12 8 ng ml 4 3 vU ABRIRU Z NERO VASTE IR IX 3 477 JV08UE 10 10 nM 10 FBS RPMI 1640 72
32. a IgE Spleen MLN Control Cu chlorophyllin Na salt Fe chlorophyllin Na salt Chlorophyll a Chlorophyll b IgE ng ml Fig 3 5 Effect of Chlorophylls and their Derivatives on IgE Producion by Rat Spleen or MLN Lymphocytes Cells were incubated for 72 hr and culture supernatant was applied to ELISA n 4 to measure IgE concentration Values in the same group without a common letter are 28 d significantly different at p 0 05 4 EE DEP OVA DEP
33. eset Con 9x4 Jp Ee X76 IgA IgM y i i Ne ho l Table 4 2 Dose dependent Effect of Ethanol Extracts of Spices on IgG Production by Mouse Spleen Lymphocytes Spice Allspice powder Onion powder Garlic Curry Clove powder Pepper black Pepper white Horseradish Cinnamon Ginger Sage powder Thyme powder Chili powder Red pepper Nutmeg Paprika powder 103 ray 1 5 1 6 1 6 1 8 1 6 1 6 1 6 Tz Ll hag eZ l z il ATOZ 1 08 no IN 1 5 1 2 1 4 1 09 1 09 1 08 1 0 1 06 1 05 1 04 0 96 0 97 0 97 1 02 107 1 3 1 2 1 3 1 4 T2 1 04 2 0 94 1 09 1 0 1 0 1 01 1 02 0 91 0 88 0 91 10 po 1 3 1 0 1 3 1 08 0 94 1 2 1 06 0 95 1 05 1 0 9197 0 97 0 95 0 IgG concentration of the control culture was 2 2 ng ml Relative IgG content 10 1 4 1 02 1 0 17 hal 1 08 1 0 0 98 0 91 0 88 1 03 1 02 0 94 0 94 10 ug ml 1 5 135 1 0 1 3 Le 1 02 pes L5 0 22 0 50 0 07 0 50 1 04 1 02 0 03 1 06 Spice jos Hy x TO t0 perii Allspice powder 1 7 Wr ky 1 7 5 1 9 Onion powder L5 1 5 0560 9 1 6 1 6 Garlic jum ls Uo 17 i R5 Curry 1 2 1 5 0 57 X6 17 1 09 Clove pow
34. IgE b Worden et al 31 1955 T B Pb Zn Cu Steffensen et al 1994 Boroskova er al 1993 Bumgardner et al 1993 Mehanna et al 1994 Smith et al 1996 Chlorophyll Iron Chlorophyllin Na Copper Chlorophyllin Na Chlorophyll a R CH3 Chlorophyll b R CHO Fig 3 8 Structure of Chlorophyll and its Derivatives Fe
35. IgE T B IgE IgE mRNA Pg RT PCR z E ee uii 61 ee WEHI 279 Ps IgM IgE prd T Ge Alt et al 1982 Gg IgM IgE Pg lgE Gg Pg Epstein Barr
36. IgE Helbling et al 1993 Niinimaki et al 1995 BIND Ae VAGA S E theo tL OU gAVEA IB URL wo AYR aT RUM FOI AED i OCI eG EH IRT o f DNA Randerath et al 1993 N o t butoxycarbonyl fructoselysined3 XQ N a t butoxycarbonyl arginine Mpento
37. 20 FBS RPMI 1640 2 WEHI 279 Table 2 2 IgM IgG WEHI 279 IgM IgO Double producer IgM IgG 0 Double producer Table 2 2 Immunoglobulin Concentration in the Culture Supernatant of WEHI 279 Clones Clone No IgM IgG J Clone No IgM IgG Clone No IgM IgG ug ml ng ml ug ml ng ml ug ml ng ml 1 T0 19 7 24 gu Wee 47 99 11 4 2 98 140 25 91 12 1 48 8 9 59 3 9 8 13 1 26 9 6 10 2 49 10 1 15 8 4 AS 70 27 92 97 50 10 1 9 1 5 97 133 28 8 9 10 5 51 96 80 6 96 11 0 29 oW 1h 52 6 9 49 7 98 14 1 30 92 88 8 TOL TSO 31 94 11 7 9 ge 45 5 32 9 7 133 10 96 13 6 33 9 5 201 11 98 145 34 9 6 156 12 9
38. Fig 2 5 KI WEHI 279 Pg Pu Table 2 1 Sequences of Primers Used in the Experiment Amplified RNA Primer Sequence Size of Amplified Product Position and Reference Pu Jug 5 TGGACTACTGGGGTCAAGG 3 no numbered Gough et al 1981 Cu 5 CATTGGGATTCATCTCTGCG 3 439 458 Kawakami etal 1980 Pv1 Jug 5 TGGACTACTGGGGTCAAGG 3 no numbered Gough et al 1981 Cy1 5 GACAGATGATACTTCTGGGA 3 884 903 Honjo et al 1979 Py2b Jug 5 TGGACT ACT GGGGT CAAGG 3 no numbered Gough et al 1981 Cy2b 5 TCCACCCTCGAGGTTAGGAG 3 954 973 Kataoka et al 1979 Py3 Jug 5 TGGACTACTGGGGTCAAGG 3 no numbered Gough et a 1981 Cy3 5 TCCACCCAAGATGTTACCAG 3 2811 2830 Wels et al 1984 Pe Ju4 5 TGGACTACTGGGGTCAAQGG 3 no numbered Gough et al 1981 Ce2 5 AGCGATGAATGGAGTAGC 3 991 1008 Ishida et al 1982 Gt lg 5 ACTAGAGAT TCACAACG 3 771 778 Gerondakis 1990 Ce2 5 AGCGATGAATGGAGT AGC 3 991 1008 Ishida et al 1982 Gy ly1 5 CGAGAAGCCTGAGGAATGTG 3 351 370 Honjo et al 1979 Cy1 5 GACAGATGATACTTCTGGGA 3 991 1008 Ishida et al 1982 p Actin sense 5 TGGAATCCTGTGGCATCCATGAAAC 3 antisense 5 TAAAACGCAGCTCAGTAACAGTCCG 3 382 bp 392 bp 419 bp 401 bp 365 bp 423 bp 418 bp 348 bp 2 3 4 5 Fig
39. FCF GO 9 Eig 3 McumL Sab ie IgG IgM 10 uM 10 nM IgE FCF FCF 10 10 uM IgE PCR PTS deg gem mp peraiyrgossg OTE Mee 10 uM Zee th FCE C 1068 10 MM IgE MLN 1 uM IgE 10 nuM IgG IgM Fig 3 4 MLN
40. Curcuma longa Lietal 1993 Sui et al 1993 Yasuni er al 1993 Soni etal 1992 Fujiyama Fujiwara et al 1992 Ammon et al 1993 Srivastava et al 1995 Huang etal 1991 Kakar et al 1994 Lu et al 1994 TPA Yasukawa et al 1994 Ere A Adegoke et al 1994 2
41. 2 Bl T B WEHI 279 B qe 0 8 1 80 IgE IgG 0 6 a S 60 A E f Sus 2 40 E ou a BEN E i 7 SS 0 2 20 a 0 0 ba 0 2 4 6 8 10 0 2 4 6 8 10 Time day Time day IgM ng ml Time day Fig 5 4 Time courses of Immunoglobulin Production by WEHI 279 Cells Cells were cultured with PBS O 1 uM of RB 6 10 uM of RB BD 100 uM of RB A 1 mM of RB A for various periods and the culture supernatant was collected to measure the Ig concentrations by ELISA The results indicated here were the mean SE n3 54 F WEHI 279 100 uM 90 1 mM 0
42. 100 uM Fig S AI 98 Wie Foi 0 10 0M OO EE SAR A eee SR HERD IgG 4 60 ng ml IgM IgG 4 300 ng ml 100 uM IgG IgM 8 UT a 100 utM IgE ie PETOS OD 10uMMW 100 uM IgE 0 1 uM IgE WEHI 279 IgE IgE WEHI 279 24
43. 37 13 A c4I5nin CURAE RE eave Liz 3 WEHI 279 1x10 cells ml 1 2 3 4 6 8 Fig 2 2 3 WEHI 279 6 i Es a s PM 3 S x a 1 Cell number Time day Fig 2 2 Growth Curve of WEHI 279 Cells Values are means SE n 2e WEHI 279 WEHI 279 1x10 cells ml 1 2 3 4 6 8 IgG IgM ng ml IgE optical density OD 400 0 20 Ty 300 0 15 T E 9 hm she _ Z O Q o 2 A Oo 100 7 LU D 0 05 OF 0 00 Time day Fig 2 3 Time course of Immunoglobulin Accumulation in Culture Supernatan
44. LTB Ford Hutchinson et al 1980 CD23 B IgE Dugas et al 1990 Yamaoka etal 1989 ro EX e 7 te 4 FH AEH 2e Bao o RAD A LL SUE A SB x 7 X 4A OMY ZI UF evuvflm dojli c ch vellus caoWwEHI 279ftllfu 2 WEHI 279 SS BD Mim
45. 6 10 gday 800 nuM 400 IgE IgG IgM 1 mM 100 uM IgG lgM 10 uuM Zr d avy el mg IgM 1 nM IgM Yamada et al 1989 1990a 1990b 1991 Maeda et al 1990 FEBZRR2 2
46. B 2C4 F3 DEP DEP Ah Pe Tsien et al 1997 Pe WEHI 279 Gg Gy1 2 IgM IlgE IgG1 IgM IgE IgG
47. HRP IgG F ab Zymed HRP IgM Fab Zymed HRP lIgA Zymed IgE IgE EIU HRP Prozyme San Leandro CA 2 9 Brown Norway MF iafe Lim etal 1994a MLN RPMI 1640 HKA K KOR Huhu cds KOMLNG 3 0 4 LC Lympholyte Rat Cedarlane Hornby Canada
48. WEHI 279 TEER OBE IZOVW CHA LK 528i 1 8 Balb c RPMI 1640 HKN RR finu cz TO OS UT 7s KH Lympholyte Mouse Cedarlane Hornby Canada Hv C HIE 10 ntergen Purchase NY RPMI 1640 2 x 10 cells ml 5 CO 72 WEHI 279 WEHI 279 10 FBS GIBCO BRL Grand Island NY RPMI 1640 1 x 10 cells ml 5 CO IgG IgG Zymed San Fr
49. 10 v PBS TRIlzol GIBCO BRL f RNA RNA 75 10 ug RNA United States Biochemical Cleveland OH RNA DNA cDNA 0 4 units Taq Fermentas Vilnius Lithuania 2 5 mM 200 iM 10 nM cDNA Gene Amp PCR system 2400 Perkin Elmer t Foster City CA Fig 2 1 DNA Denaturation Primer Extension 94 C 0 5 min Primer Annealing 72 C 1min 55 1 min RE 35 Cycles Fig 2 1 Temperatures
50. rro WEHI 279 WEHI 279 B IgM x Warner etal 1979 Gutman et al 1981 D KE WEHI 279 55 2 Bh 1 FITC IgE FITC IgG FITC IgM Southern Biotec
51. Block Ace IgG IgG F ab Cappel West Chester PA IgE IgE Fc 7S Nordic Immunology Tilburg The 18 A Netherlands IgM IgM Fab Zymed San Francisco CA HRP IgG Fab Cappel HRP IgM Fab Cappel IgE IgE Zymed HRP DAKOPATTS Copenhagen Denmark 2 9 Brown Norway CET Zdi amp m du MF
52. 1I IL 4 IL IFN c IFN y Elson er al 1994 Gauchat et al 1991 Ochel et al 1991 Pene et al 1988 Rousset et al 1991 Lim etal 1994a 1994b Yamada et al 1993 Wy Aidt mM MLN IgE lgA IgG lgM Lim et al 1994b A ConA IgE PWM Lim er al 1994b 1gE acce SA sug vp OVID eS Se OY le see ER Plaut et al 1993 LT Dahlen er al 1986
53. 37 1 4 19 4 3 37C 1 IgE 37C 1 37C 1 37C 13 415nm 4 Student s test Duncan Duncan 1955 HIM 1 Table 3 1 1 mM IgE 1 mM IgE IgE
54. MLN MLN Fig 3 2 IgG IgM IgE 1 5 10 wM IgE 4 HCFE
55. Muranaka et al 1986 Takafuji et al 1987 1989 Takenaka et al 1995 Savage etal 1991 TRU 005 Dd CWA 8 Nish et al
56. MLN IgE IgA IgG IgM Lim etal 1994b A ConA IgE PWM Lim et al 1994b 2 1 SPORTS PEDI eg day KEA PIENE EL MH poe UV RE RT 5N OO RIPC 3 L 3 3 5 L Aldrich St Louis MO B Janssen Chimica Brussels Belgium 0 05 Tween 20 PBS TPBS Bloc
57. WEHI 279 B WEHI 279 lsM Gutman et al 1981 WEHI 279 B Sibley et al 1980 1981 Paige et al 1982 Gon Ole wegen wal yer AE CLA RAV HVED Y IRRI SERERA L Hae bia IgE IgG IgM WEBEHI 279 100 uM 24 OL
58. Takafuji et al 1987 1989 Takenaka et al 1995 L L YL VX BFO 1 IgE Metcalfe 1991 A IgE Lim et al 1994 Lim er al 1994 1 10 uM IgE 0 1 2 3 nM Curcuma onga
59. 1991 Tarlo et al 1993 Weber 1993 Wuthrich er al 1993 JG 1 3525 99 e ZA JC oV TI AM EX OO 4 1 Metcalfe 1991 lgE lgG 1L 4 IL IFN 46 a IFN y Elson et al 1994 Gauchat et al 1991 Ochel et al 1991 Pene et al 1988 Rousset et al 1991 Lim etal 1994a 1994b Yamada et al 1993 IERI mM
60. 2 5 Expression of Productive mRNAs in WEHI 279 Cells Each separated mRNA from WEHI 279 cells was amplified by RT PCR Each lane was indicated as Lane 1 u mRNA Pu Lane 2 Py3 Lane 3 Pyl Lane 4 Py2b Lane Sub Fig 2 6 Gg Gy1 WEHI 279 IlsM IgG1 IgE ACE Fig 2 6 Expression of Germ line mRNAs in WEHI 279 Cells Each separated MRNA from WEHI 279 cells was amplified by RT PCR Each lane was indicated as Lane 1 yl mRNA Gyl Lane 2 Ge WEHI 279 IgM IlsG IgE 0 96 1 0 8 96 10 1 1
61. Fig 2 4 IgM IgsG IgE WEHI 279 IgM IgG IgE FITC anti lgG FITC anti IgM Fig 2 4 Expression of Surface Immunoglobulin on WEHI 279 Cells surface Igs were detected by flow cytometer WEHI 279 mRNA WEHI 279 1x10 cells 2 2 3 mRNA umRNA Pu Py1 Py2b Py3 Ps IgM IgE s Ge IgM IgG1 Gy1 RT PCR B actin cDNA Table 2 1
62. IL 30 ON vite H CH DIET TCLPS ZIMU ZZ PRN VY OSEROI SEE EIS HH A IVS 23 MLN IgE Lim et al 1994b 1995 MLN Morales et al 1995 TY ONES IAB dE AVA IgG IgM SO Na inthe Omen Os N C4H3 CH SO Na SO Brilliant Blue FCF Fast Green FCF Fig 3 7 Structure of Triphenylmethane Colorings 2 VP PAE CDOT VAI OA FERIA hAU YFCF brunu UY FRU AI RY LER CIS p M MLN 10 uMEI EO RUE CHRIS EK OMLNY z 3EROIgE
63. h Hi PBS PBS Fig 5 5 100 IM IgE 10 uM IgE 1 10 LM IgG 100 uM lg 100 uM WEHI 279 IgE lgE Extracellular Immunoglobulin Level Culture Supernatant Control HB 1 M RB 109M RB 100 M OO OBS 15 5 pe xo Wwe g b20 4050 80 100 IgE A415 IgG ng ml IgM ng ml Intracellular Immunoglobulin Level Control HB 1 M RB 109M RB 1000M 00 Q2 OM 96 QB 0 5 10 15 QO 20 40 60 80 106 IgE A415 IgG ng ml IgM ng ml Fig 5 5 Effect of RB on Intracellular and Extracellular Immunoglobulin Levels of WEHI 279 Cells Cells were cultured for 24 h in the absence or presen
64. 0 01 LOF 20 X0 1 01 0 02 1 12 0 04 1 1820 04 03620 4 99 Lo 1 12 0 07 0 43 0 00 1 0730 02 0 94 0 03 9 OG Ig concentrations of the control culture were 6 81 0 51 ng ml for IgE 30 5 0 92 ng ml for IgG and 78 2 2 3 ng ml for IgM Values are the mean SD of 4 cultures p lt 0 05 p lt 0 01 and p lt 0 001 compared with the control 2 Dy PRU ZZ NEKOAA AFE E IC X d 3bEZKASEXSAG ORE WRZ Dy PRY LY ASBRO PTI EEZ RIE 3E IKTATVEX BR fh 45 00 EE A UPC OIE Table 3 2i E RIC EOI Peery ay Be DIT 1 100 uM IgE 100 uM IgG IgM 1 10 uM CIgMIBE ze ESL AI IgM 100 unuM IgG Table 3 2 Dose dependent Effect of Water insoluble Colorings on Immunoglobulin Production by Rat Spleen Lymphocytes Coloring Concentration Relative Ig content uM gk IgG IgM Control 0 1 00 0 04 1 00 0 02 1 00 0 01 Gardenia yellow 0 54 0 11 1 04 0 01 1 24 0 04 10 67 41 9 1 04 0 04 1 1620 04 100
65. 0 83 0 06 0 9030 03 AR EOS Laccaic acid 0 4640 09 1 06 0 03 Zoe Oar ae 10 0 45 0 07 1 1640 07 SI RN OF 100 0 61 0 09 1 40 0 12 L620 Lms Bixin 0 5440 04 1 04 0 02 EIT ai 10 Q0 5620 067 1 03 0 03 1 252020458 100 0 80 0 04 0 96 0 03 1320 05 Curcumin 0 92x0 11 1 0530402 1 58 80 058 55 10 0 86 0 11 1 14 0 10 290r Gr 100 0 8630 10 Ode OS Q740002 The Ig concentrations of the control culture were 6 45 0 25 ng ml for IgE 24 14 0 52 ng ml for IgG and 64 70 95 ng ml for IgM Values are the meanrSD of 4 cultures p lt 0 05 p 0 01 and p 0 001 compared with the control 3 MLN C NENH Jo Pe ee E 2 2 GE 72 PBS
66. 1 11 3 35 98 10 9 13 99 14 6 36 96 12 9 14 9 0 157 37 5 11 0 15 10 2 14 2 38 96 129 16 94 120 39 95 14 7 17 92 10 0 40 95 81 18 96 18 0 41 9 7 11 7 19 79 80 42 10 0 12 6 20 97 140 43 9 8 149 21 93 125 44 76 70 22 90 95 45 amp 6 ENI 23 94 11 9 46 95 9 SRABD ER MLN TI Armitage et al 1993 Collins et al 1995 Ling et al 1995 B B WEHI 279 AX WEHI 279 Walter and Eliza Hall Institute WEHI Melbourne Australia WEHI 279 B IgM Ka
67. 1993 Wuhrich 1993 Nish etal 1991 Adegoke et al 1994 URL EERO Zheng et al 1992 Rao et al 1995 Cgpsjcun annum Higashimoto et al 1993 Azizan et al 1995 Sastre et al 1996 Cooper et al 1996 Kanerva et al 1996 aniseed Fraj etal 1996
68. GTAGC 3 991 1008 Ishida et al 1982 B Actin sense 5 TGGAATCCTGTGGCATCCATGAAAC 3 348 bp antisense 5 TAAAACGCAGCTCAGT AACAGTCCG 3 DNA Denaturation 94 C 0 5 min Primer Extension Primer Annealing 72 C 1 min 55 C 1 min eM 35 Cycles EHP p Fig 5 6 RT PCR Schedule B actin cDNA Table 5 1 Fig 5 7 1 100 LM WEHI 279 Pg WEHI 279 Gg Gzg WEHI 279 Pg IgE Ge IgM Ig E LaneNo 1 2 3 4 Fig 5 7 Effect of RB on the Expression of Pe or Ge of WEHI 279 Cells Cells were cultured for 72h in the presence or absence of RB at each concentration and RNA was extracted from the cells to apply to RT PCR Eac
69. PS on IgE Production by WEHI 279 Cells Cultured with or without RB Cells were incubated with each concentration of Rose Bengal for 48 h and culture supernatant was applied to ELISA to measure Ig concentration Hatched or open column indicates the culture with or without LPS 2 5 ug ml respectively Values in the same group without a common letter are significantly different at p 0 05 n 4 4 EE 3 50 uM IgE IgG IgM 4 4 IgE 4 I
70. ancisco CA IgE IgE EIU Brussels Belgium IgM IgM F ab Zymed 34 IgA lgA Zymed HRP IgG F ab Zymed HRP IgM Fab Zymed HRP AlgA Zymed IgE IgE EIU HRP Prozyme San Leandro CA 2 pl c MU UAE EE NIME Ztt 7 BH EM Gea YE Yaa ey eA BPA es JEEP 3 S t tg 77 VASO AS 1g 10 ml 30 10 g ml 100 ug ml 0 001 pg
71. and Reaction Periods Used During Amplification Cycles PCR 2 Sawady Technology DNA Kodak Digital Science Eastman Kodak t Rochester NY Cf mw LU t 4 Yamada etal 1993 IgG IsM IgE 96 37 1 TPBS 3 37 1 4 C 4 3 37 1 IgE 37C 1 37 1
72. ce of RB and the Ig concentration in the culture supernatant were determined for result of A or were lysed by sonication to determine the Ig concentration in the cells for result of B by ELISA Values in the same group without a common letter are significantly different at p lt 0 05 n 3 3 WEHI 279 IgE WEHI 279 72 RNA C gmRNA Pe IgM IgE 0 s Ge iAle Fig 5 60 RT PCR Table 5 1 Primer Sequences Wsed iin the Experiment Amplified RNA Primer Sequence Size of Amplified Product Position and Reference Pe J 9 TGGACTACTGGGGTCAAGG 3 365 bp no numbered Gough et al 1981 Ce 2 5 AGCGATGAATGGAGTAGC 3 991 1008 Ishida et al 1982 Ge le 5 ACTAGAGATTCACAACG 3 423 bp 771 778 Gerondakis 1990 Ce2 5 AGCGATGAATGGA
73. der Ls 1 09 04 15 1 2 1 09 Pepper black 096 109 0 32 1 2 0 96 0 96 Pepper white 109 0 83 0 44 1 3 0 83 0 96 Horseradish 1 3 Jy 0 44 1 09 1 09 0 96 Cinnamon 0 96 1 2 0 86 1 4 12 0 61 Ginger 1 2 1 5 1 0 142 1 01 0 77 Sage powder 0 94 1 2 1 1 1t 1 9 0 52 Thyme powder 1 060 0 97 Ig Tj 0 98 0 63 Chili powder 2 1 04 1 02 1 3 1 2 1 06 Red pepper 1 1 MEL 0 94 0 91 0 96 12 Nutmeg 03 4096 WWF LOL 097 0359 Paprika powder 1 09 1 05 1 01 1 09 1 2 fs Table 4 3 Dose dependent Effect of Ethanol Extracts of Spices on gA Production by Mouse Spleen Lymphocytes Relative IgA content IgA concentration of the control culture was 1 1 ng ml 218 WEHI 279 B B WEHI 279 WEHI 279 IgM IgG Table 4 4 IgM I 39 b SV ZG
74. f F r f a 3 LI I P Li f Li e J f BASIE Ef 1998 As 1 rE 425 WEHI 279 Tae MMe WES NT SP n m mese eese 5 BS FE klik Mu 45 TOCRROBUAPE ESIBIERBE esses 3 3 BIE Less 4 6 HOR LS ANS DV Orme BTR i 7 6 WM 9 a Ce al eS er ear o a cl t 101 1 f a RNO SAT ROA vixi ROF CAS CAMEL fEA 5I ovv TILK KAEASHEA T O RAR E CHAT h ARAT ETE BUILT 9 Niebauer et al 1995 Loft et al 1996 Tarlo et al 1993 Weber 1993 Wuhrich 1993 Nish et al 1991
75. gE 4480 RET IAS m Ze Ito et al 1986 gt Br gt H Legrum 1991 HAAR UAT MLN IgE
76. globulin Production by Rat Spleen Lymphocytes Lymphocytes were cultured in the presence of 100 uM xanthene dyes for 6 h to measure the IgE content and for 72 h to measure the IgG and IgM contents Each result is the mean SE n 4 and 2 lvalues without a common letter are significantly different at p 0 05 Fig 5 2 Fig 5 3 IgE IgG IgM 4 4 IgE 4 A Re RL 4 4 IgE
77. h lane indicates Lane 1 Control PBS Lane 2 1 uM RB Lane3 10 uM RB Lane 4 100 uM RB LPS B Gronowicz et al 1974 WEHI 279 LPS WEHLI 279 LPS 2 5 ug ml 0 100 uM 48 Fig 5 8 LPS WEHI 279 IgE IgE 9 0 10uMM LPS WEHI 279 IgG IgM 100 uM IgG IgM LPS 3 V 58 B Control RB 1 uM RB 10 uM RB 100 uM 20 30 ng ml ng ml Fig 5 8 Effect of L
78. hnology Associates Birmingham AL 0 05 Tween 20 PBS TPBS Block Ace lgG IgG Zymedft Sanfrancisco CA IgE IgE EIU Brussels Belgium IgM IgM Fab Zymed T IgA IgA Zymed CHRP IgG F ab Zymed HRP IgM F ab Zymed HRP IgA Zymed IgE IgE EIU HRP Prozyme San Leandro CA 2 WEHI 279
79. k Ace IgG IgG Fab Cappel West Chester PA IgE IgEFc 7S Nordic Immunology Tilburg Netherlands lIgM IgM F ab Zymed San Francisco CA CARP 47 LL IgG Fab Cappel k OHRPR RV 2 IgM F ab Cappel IgE IgE Zymed HRP DAKOPATTS Copenhagen Denmark IgG IgG Zymed IgE lIgE EIU Brussels Belgium IgM IgM Fab Zymed IgA IgA Zymed
80. ml Dose response 3 Yamada etal 1993 IgG IgM IgE 96 37C 1 TPBS 3 37C 1 4 C 4 3 37C 1 IgE 37 1 37 1 37 13 i 415nm y 3 1
81. rings such as triphenylmethane dyes brilliant blue FCF and fast green FCF indigoid dye indigocarmine water soluble annatto and sodium Cu chlorophyllin at around 10 to 10 uM for 72 hr and the culture supernatant was collected to evaluate Ig concentration 5 MLN MLN V4 ded Uv ZUG Ud POAT AYE AZ BET 4 aa 2 b Fig 3 5 100 uM IgE b MLN IgE
82. sidine Oya et al 1997 LEDT FF WEHI 279 T 0 1 5 2 30 nM 5 1 40
83. t of WEHI 279 Cells Cells were cultured for various days and Ig concentration in the culture supernatant was measured by ELISA IgG O and IgM concentrations were indicated as ng ml and that of IgE as OD Optical density Fig 2 3 IgG IgM 4 IgE 1 2 IgE WEHI 279 4 HUAI Aid L lt RE E WEHI 279 IgM Gutman etal 1981 WEHI 279 WEHI 279 10 lx10 cells ml 2 2 2

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