1209 URACIL IN TRANSFER FACTOR
SIR,-The biological and biochemical nature of leucocyte transfer factor (T.F.) is unknown, but it was originally thought to represent an informational molecule with immunological specificity.’ For chemical structure, polynucleotides and polypeptides have been considered, and several groups have demonstrated nucleotides in fractions obtained by chromatography on ’Sephadex G-25’ and ’G-10’ columns.2Characin these studies T.F. activity has always been found in fractions, which have adsorbed to the gel and eluted later
teristically,
than small inert molecules, such
as
NaCl
1(Kav =Vo-Vt >
1).
Adsorption to sephadex G-25 or G-10 occurs with molecules containing aromatic structures which have a -r electron and are small enough to enter the inner volume of the column (less than 700 daltons for sephadex G-10). For these reasons, it would be unlikely that fractions adsorbing to sephadex G-10 gel could contain molecules large enough to carry immunological information. We obtained nine main fractions from leucocyte dialysates with chromatography on sephadex G-10 columns and studied the therapeutic value and the capacity of these fractions to induce skin reactivity or strengthen previously weak skin reactions to natural antigens, such as P.P.D. One of the fractions, designated via and having a Kav value of 1.68, was consistently found to have T.F. activity, but fractions I, III, and vn were active occasionally. Repeated injections of this fraction at a dose of 15 fig weekly had a clear therapeutic effect (see accompanying table). Characteristically, the increased skin
EFFECT OF TREATMENT WITH FRACTION via ON THE CLINICAL PICTURE AND ON SKIN REACTIVITY TO PPD
,
Thin-layer chromatography of fractions sephadex G-10 column.
These fractions represent material which adsorbed to the gel The plate was developed in chloroform/methanol/17% ammonium hydroxide (40/40/20) and photographed in short l1.V. light. U uracil, Hx = hypoxanthine, Tyr tyrosine, R.N.A. ribonucleic acid. =
=
We suggest
least in patients with impaired cellular of T.F. is based on non-specific stimulamechanism of this stimulation, the possible relation of T.F. to the thymic hormone, T.H.F., is of interest. T.H.F. has been shown to increase the level of cyclic adenosine monophosphate (CA.M.P.) in spleen cells,. while one the active fraction of T.F. increased the level Of CG.M.P. in blood monocytes.’ As to the substances which we have identified in T.F., hypoxanthine is known to potentiate the action of CA.M.P. on the cells of adrenal cortex.6 It is possible that both the thymic hormones and T.F. are involved in different phases of the same reaction chain, resulting in the activation of cells involved in the expression of cellular immune response.
that,
Finland.
associated with- an alteration in the blast-transformation test. Further characterisation of the different fractions was performed with thin-layer chromatography. The accompanying figure demonstrates spots absorbing u.v. light at 254 nm in fractions iv - IX, all with a Kav value greater than 1.0. The material in the only spot seen in fraction via was eluted from the plate and its T.F. activity was tested in patients with generalised sarcoidosis having a weak or missing skin reactivity to P.P.D., oidiomycin, and streptokinase/streptodornase. 30 ug of eluate was effective in inducing or strengthening skin reactivity. Based on stain reactions, Rf values, hydrolysis experiments, and I.R. spectrometry, the two spots in fraction v were identified as tyrosine and a small R.N.A. molecule containing three mononucleotides and the spot in fraction vi) as hypoxanthine. The only spot seen in fraction via was shown to contain uracil as the major component. It thus seems possible that the claims of the presence of polynucleotides in active T.F. fraction are based on analysis of impure preparations,2and that some of the effects of T.F. may be mediated by uracil or by substances closely related to it.
reactivity to test antigens was not
1. Lawrence, H. S. Adv. Immun. 1969, 11, 195. 2 Baram, P., Yuan, L., Mosko, M. M.J Immun. 1966, 104, 769 3. Gottlieb, A. A., Foster, L. G., Waldman, S. R. Lancet, 1973, ii, 822.
=
at
immunity, the effect tion. In defining the
University of Tampere, 33520 Tampere 52,
T.u.=tuberculin unit.
a
(Kav>1.0).
Institute of Biomedical School of Medicine,
Grading of skin tests: 0=100 T.U. negative, 1=100 T.u. positive, 2=10 T.U. positive, 3=1 T.u. positive, 4=0-1 T.u. positive.
VI - IX obtained from
Sciences,
K. J. E. KROHN A. UOTILA P. GRÖHN J. VÄISÄNEN
BLOOD-TRANSFUSION IN RHEUMATOID ARTHRITIS 70 SIR,-A aged was diagnosed as having seropositive, anodular rheumatoid arthritis (R.A.) in 1971, and since then he had been on ibuprofen 400 mg three times a day, enteric coated aspirin 650 mg four times a day, indomethacin capsules 50 mg twice daily, and an indomethacin 100 mg suppository at night. As there was little change in his symptoms, he was started on sodium aurothiomalate from January 1972. However, in spite of these injections he continued to have exacerbations of his arthritis. In March 1975, severe joint pain developed and he took several aspirin tablets. He was later admitted to Black Notley Hospital with haematemesis and melsena. On admission he was pale and had R.A. changes in both hands, wrists, and elbows. The joints were warm and very painful. Blood-transfusion was started and at one time his haemoglobin dropped to 6.7 g/dl. He received 4 units of B-negative whole blood during the next 3 days. He improved dramatically and could make a complete fist and move all the involved joints without experiencing pain. His morning stiffness also disappeared (before admission it had lasted 4-6 hours). Unfortunately there were no previous erythrocyte-sedimentation-rate results for comparison. At present the patient is taking an indomethacin suppositorv man
4. 5. 6.
Kook, A. J., Trainin, N.J. Immun. 1975, 115, 8. Kirkpatrick, C. H. J. Allergy clin. Immun. 1975, 55, 411. Vapaatalo, H., Bieck, P., Westermann, E. Nannyn-Schmiedeberg’s Pharmak. 1972, 275, 435.
Arch
SIR,-The biological and biochemical nature of leucocyte transfer factor (T.F.) is unknown, but it was originally thought to represent an informational molecule with immunological specificity.’ For chemical structure, polynucleotides and polypeptides have been considered, and several groups have demonstrated nucleotides in fractions obtained by chromatography on ’Sephadex G-25’ and ’G-10’ columns.2Characin these studies T.F. activity has always been found in fractions, which have adsorbed to the gel and eluted later
teristically,
than small inert molecules, such
as
NaCl
1(Kav =Vo-Vt >
1).
Adsorption to sephadex G-25 or G-10 occurs with molecules containing aromatic structures which have a -r electron and are small enough to enter the inner volume of the column (less than 700 daltons for sephadex G-10). For these reasons, it would be unlikely that fractions adsorbing to sephadex G-10 gel could contain molecules large enough to carry immunological information. We obtained nine main fractions from leucocyte dialysates with chromatography on sephadex G-10 columns and studied the therapeutic value and the capacity of these fractions to induce skin reactivity or strengthen previously weak skin reactions to natural antigens, such as P.P.D. One of the fractions, designated via and having a Kav value of 1.68, was consistently found to have T.F. activity, but fractions I, III, and vn were active occasionally. Repeated injections of this fraction at a dose of 15 fig weekly had a clear therapeutic effect (see accompanying table). Characteristically, the increased skin
EFFECT OF TREATMENT WITH FRACTION via ON THE CLINICAL PICTURE AND ON SKIN REACTIVITY TO PPD
,
Thin-layer chromatography of fractions sephadex G-10 column.
These fractions represent material which adsorbed to the gel The plate was developed in chloroform/methanol/17% ammonium hydroxide (40/40/20) and photographed in short l1.V. light. U uracil, Hx = hypoxanthine, Tyr tyrosine, R.N.A. ribonucleic acid. =
=
We suggest
least in patients with impaired cellular of T.F. is based on non-specific stimulamechanism of this stimulation, the possible relation of T.F. to the thymic hormone, T.H.F., is of interest. T.H.F. has been shown to increase the level of cyclic adenosine monophosphate (CA.M.P.) in spleen cells,. while one the active fraction of T.F. increased the level Of CG.M.P. in blood monocytes.’ As to the substances which we have identified in T.F., hypoxanthine is known to potentiate the action of CA.M.P. on the cells of adrenal cortex.6 It is possible that both the thymic hormones and T.F. are involved in different phases of the same reaction chain, resulting in the activation of cells involved in the expression of cellular immune response.
that,
Finland.
associated with- an alteration in the blast-transformation test. Further characterisation of the different fractions was performed with thin-layer chromatography. The accompanying figure demonstrates spots absorbing u.v. light at 254 nm in fractions iv - IX, all with a Kav value greater than 1.0. The material in the only spot seen in fraction via was eluted from the plate and its T.F. activity was tested in patients with generalised sarcoidosis having a weak or missing skin reactivity to P.P.D., oidiomycin, and streptokinase/streptodornase. 30 ug of eluate was effective in inducing or strengthening skin reactivity. Based on stain reactions, Rf values, hydrolysis experiments, and I.R. spectrometry, the two spots in fraction v were identified as tyrosine and a small R.N.A. molecule containing three mononucleotides and the spot in fraction vi) as hypoxanthine. The only spot seen in fraction via was shown to contain uracil as the major component. It thus seems possible that the claims of the presence of polynucleotides in active T.F. fraction are based on analysis of impure preparations,2and that some of the effects of T.F. may be mediated by uracil or by substances closely related to it.
reactivity to test antigens was not
1. Lawrence, H. S. Adv. Immun. 1969, 11, 195. 2 Baram, P., Yuan, L., Mosko, M. M.J Immun. 1966, 104, 769 3. Gottlieb, A. A., Foster, L. G., Waldman, S. R. Lancet, 1973, ii, 822.
=
at
immunity, the effect tion. In defining the
University of Tampere, 33520 Tampere 52,
T.u.=tuberculin unit.
a
(Kav>1.0).
Institute of Biomedical School of Medicine,
Grading of skin tests: 0=100 T.U. negative, 1=100 T.u. positive, 2=10 T.U. positive, 3=1 T.u. positive, 4=0-1 T.u. positive.
VI - IX obtained from
Sciences,
K. J. E. KROHN A. UOTILA P. GRÖHN J. VÄISÄNEN
BLOOD-TRANSFUSION IN RHEUMATOID ARTHRITIS 70 SIR,-A aged was diagnosed as having seropositive, anodular rheumatoid arthritis (R.A.) in 1971, and since then he had been on ibuprofen 400 mg three times a day, enteric coated aspirin 650 mg four times a day, indomethacin capsules 50 mg twice daily, and an indomethacin 100 mg suppository at night. As there was little change in his symptoms, he was started on sodium aurothiomalate from January 1972. However, in spite of these injections he continued to have exacerbations of his arthritis. In March 1975, severe joint pain developed and he took several aspirin tablets. He was later admitted to Black Notley Hospital with haematemesis and melsena. On admission he was pale and had R.A. changes in both hands, wrists, and elbows. The joints were warm and very painful. Blood-transfusion was started and at one time his haemoglobin dropped to 6.7 g/dl. He received 4 units of B-negative whole blood during the next 3 days. He improved dramatically and could make a complete fist and move all the involved joints without experiencing pain. His morning stiffness also disappeared (before admission it had lasted 4-6 hours). Unfortunately there were no previous erythrocyte-sedimentation-rate results for comparison. At present the patient is taking an indomethacin suppositorv man
4. 5. 6.
Kook, A. J., Trainin, N.J. Immun. 1975, 115, 8. Kirkpatrick, C. H. J. Allergy clin. Immun. 1975, 55, 411. Vapaatalo, H., Bieck, P., Westermann, E. Nannyn-Schmiedeberg’s Pharmak. 1972, 275, 435.
Arch
