GB2213056A - Agents for removing immunodeficiency virus and related compounds - Google Patents

Abstract

Agents for removing human immunodeficiency virus and its related compounds are manufactured by the use of a solid substance having a weakly acidic or weakly alkaline surface, e.g. silica-alumina or a cation-exchange resin of COOH- or SO3 H-type. HIV is preferably removed from the patients blood by an extracorporeal blood circulation system using a column packed with the absorbent.

Description

AGENTS FOR REMOVING HUMAN IMMUNODEFICIENCY VIRUS AND RELATED COMPOUNDS This invention relates to removal of human immunodeficiency virus (hereinafter abbreviated as HIV) and related compounds from the body fluid of patients through adsorption.

HIV has affinity for human lymphocytes, particularly for helper T lymphocytes, and tends to invade these lymphocytes and destroy them, thus lowering the function of cell-mediated immunity. As a result, the patients often suffer from opportunistic infection or malignant tumour, which is usually fatal.

Research is actively under way to find effective remedies for infections caused by HIV. For example, attempts have been made to prepare an effective vaccine, but the obstruction is difficulty in setting the correct antigen (the target of the vaccine) because HIV gene is likely to undergo mutation. Administration of Suramin, HPA and AZT was also attempted based on the concept that a specific inhibitor against its reverse transcriptase could retard the settlement of viral genome. However, no satisfactory result has been obtained, with only their side effects being reported. It is conceivable that antiviral agents like interferons could retard the assembly of retroviruses; actually, however, no marked effect has been observed in clinical tests.An increase in the amount of immune complexes and the presence of inhibitory factors derived from interleukin-2 were observed in the plasma of HIV-infected patients, and plasmapheresis has been tried to remove these substances. But no satisfactory result has yet been obtained [ Plasma Therapy, 8, 23 (1987) ] . Many kinds of therapies have been attempted to cure the infections caused by HIV as described above, but only limited effects, if any, were exhibited by any such therapy.

Based on the information that HIV is released in large quantities from T cells into the body fluid of HIV-infected patients, particularly those in the aggravating period, the inventors considered that, if the amount of HIV or related compounds in their body fluid (particularly in the plasma) can be reduced by the extracorporeal blood circulation system, this might prevent their condition from worsening, thus prolonging their lives.

In accordance with this invention, a solid substance with a weakly acidic or weakly alkaline surface is used in the manufacture of agents for removing HIV or related compounds from the body fluid of HIV-infected patients.

The solid substances with a weakly acidic or weakly alkaline surface used in this invention are those that show a weakly acidic or weakly alkaline reaction when a pH reagent is dropped onto their surface. Weak acidity herein means a pH level in the range 2.5 to 6.9, and weak alkalinity a pH level in the range 7.4 to 10.5. Substances whose surface pH is in the range 7.0 to 7.3 (e.g. nonpolar polypropylene, quartz and cation-exchange resins of SO3 Na-type) are unable to adsorb HIV and related compounds, while substances whose surface pH is less than 2.5 (e.g. strongly acidic cationexchange resins) or higher than 10.5 (e.g. strongly basic anion-exchange resins) tend to coagulate proteins contained in the body fluid and hence fail to remove HIV or related compounds efficiently.

As examples of the solid substances with a weakly acidic or weakly alkaline surface used in this invention, there may be mentioned inorganic substances, such as calcium phosphate (e.g. hydroxyapatite), alumina, silica, silica-alumina and zirconia; and polymers bearing polar groups, such as anion-exchange resins (NH2-type), cation-exchange resins (COOH- and SO3 H-types), sulphonated, carboxy-containing or amino-containing polyolefins, sulphonated polystyrene, and insoluble sulphonated, carboxyl-containing or amino-containing polyvinyl alcohols crosslinked with an aldehyde. Each of these substances may be used alone in any desired form (e.g. as granules), or may be supported on an insoluble carrier (e.g. porous glass). Further, polyacrylic acid, heparin or dextran sulphate supported on the carrier may be used.Solid substances with a weakly acidic surface are preferred, e.g. cation-exchange resins (SOH3- and COOH-types) and silica-alumina.

Any solid substances in which weak acidity or alkalinity is exhibited only at the surface may be used for the purpose of this invention, except those that are soluble in body fluid. These solid substances may either be porous or nonporous.

Since adsorption of HIV or related compounds takes place on the surface of the solid substance, the internal structure of the solid may be of any type and may be made of different material from the surface. The agents for removing HIV or related compounds of this invention may be of any desired shape, such as powder, granules, plates and fibres, but are usually in the form of granules with a diameter in the range 0.1 to 5 mm. In addition, these granules should preferably be spherical to minimize possible damages to haematocytes, because the agent is brought into direct contact with haematocytes in some cases.

It has been unexpectedly found that treating the body fluid (e.g. blood or plasma) of a patient with an agent comprising a solid substance as defined above efficiently removes HIV and related compounds contained therein through adsorption. "HIV-related compounds" herein means metabolites produced by cells attacked by HIV, and immunosuppresive factors, i.e. substances that retard the activity of immunocompetent cells produced upon invasion of viruses.

The agent comprising a solid substance used in accordance with this invention is charged to a column for service. The column used for this purpose should preferably be provided with an inlet and an outlet designed to allow easy connection with the blood circuit, and with a filter made of, for example, polyester and set between the inlet and the agent layer, and between the outlet and the agent layer. The column may be made of glass, polyethylene, polypropylene, polycarbonate, polystyrene or polymethyl methacrylate. Of these, polypropylene and polycarbonate are preferred materials, because the column packed with the agent is sterilized (e.g. autoclave and y-ray sterilization) before use.

HIV and related compounds can be removed from the body fluid of patients by the extracorporeal blood circulation system using a packed column as described above. The following two types of extracorporeal blood circulation system may be mentioned.

(1) Blood taken from a blood vessel of a patient is forced to pass through a column packed with the agent of this invention to remove from it by adsorption HIV and related compounds, and the clarified blood from the column is returned to the blood vessel of the patient.

(2) Blood taken from a blood vessel of a patient is first separated into haemocytes and plasma, the plasma thus separated is then forced to pass through a column packed with the agent of this invention to remove from it by adsorption HIV and related compounds, the clarified blood from the column is mixed with the haemocytes separated above, and the mixture is returned to the blood vessel of the patient.

Of the two methods described above, the latter is more practical because of the smaller loss of haemocytes (for example, by adhesion of platelets and haemolysis of erythrocytes).

Such treatments as described above remove HIV and related compounds from the body fluid of patients, thereby preventing their conditions from worsening and prolonging their lives. In the aggravating period, HIV is released in large quantities from T cells as stated above, and hence this adverse tendency can be repressed if HIV is efficiently removed.

Removal of the related compounds will aslo suppress the antigen-antibody reactions caused by these compounds and allow freedom from suppressing growth of lymphocytes, thus preventing the patient's condition from worsening. It should be noted here that such treatments can also remove useful components in the body fluid. If this happens, it is preferable that supplements of these useful components be given to the patients.

The following Examples illustrate the present invention: Examples 1 to 8 and Comparative Examples 1 to 3 (1) Specifications of agents for removing HIV and related compounds Test experiments were conducted using agents of the following specifications.

Table l

No. Agent Specifcations Suriace pH Comp. Ex. Polypropylene Nonpolar powder (av. size: 0.75mm) 7.0 1 Comp. Ex. Quartz Quartz powder with its surface treated with a 7.0 2 silane coupling agent (av. size: 0.006mm) Comp. Ex. Cation-exchange SO3Na-type; IR120B of Japan Organo Co.; Granules 7.0 3 resin ( size: 0.6@m, surface aree < 0.1m/8) Example Hydroxyapatite Granules ( size: 0,45mm, surface area: 100m/g); 6.5 1 Ca/P: 1.64; calcination temp.: 550 C Example Hydroxyapatite Granules ( size: 0.45mm, surface area: 80/g); 8.5 2 Ca/P: 1.64, calcination temp.: 700 C Example Silica-alumine Product of Davison Co.; granules ( size: 0,45mm, 3.0 3 surface area: 140m/g ); Si/Al: 4.4 Example Anion-exchange NH2-type; IRA401 of Japan Organo Co.; Granules 8.5 4 resin ( size: 0.6mm, surface area < 0.1m/g) Example Cation-exchange COOH-type; IRO84 of Japan Organo Co.; Granules 3.0 resin size: 0.ii surface aree < 0.1m/8) Example Cation-exchange SO3H-type; IR120B of Japan Organo Co.; Granules 3.0 6 resin ( size: 0.6mm, surface area < 0.1m/g) Example Alumina Granules ( size: 0.45mm, surface area: 100m/g ); 5.0 7 calcination temp.: 900 C Example Alumina Granules ( size: 0.45mm, surface area: 100m/g ); 10.0 8 calcination temp.: 1200 C (Note) Measurement of surface pH: Each agent was placed on a glass plate, a pH reagent was dropped on it, and surface pH was measured by colorimetry using a pH meter of Kyowa Rikagaku Kenkyusho.

(2) Procedure for removing HIV and related compounds Culture supernatant of HIV-infected cells [HIV cell line H9 (possessed by NCI Of the United States) inoculated into sensitive cells CEM (CCRF, CEM, Flow Laboratories, ATCC No. 19, acute lymphocytic leukaemia), followed by multiplication and subculture ] was treated with each of the agents mentioned above according to the procedure detailed below.

(i) The agent (0.5 g) was immersed in 2 ml of RPMI-1640 medium, and the mixture was deaerated by heating and subjected to steam sterilization in a tightly stoppered container.

(ii! Culture supernatant of HIV-infected cells was filtered through a 0.45 um filter, and the filtrate (4 ml) was mixed with 16 ml RPMT-1640 medium.

(iii) Supernatant (2 ml) of the mixture obtained in (ii) was added to the mixture obtained in (i), and the resulting mixture was allowed to stand at room temperature for 30 minutes.

(iv) Supernatant of the mixture obtained in (iii) was filtered through a 0.45 um filter.

(v) The filtrate obtained in (iv) (0.5 ml) was added to a culture liquor [obtained by incubation of cord blood lymphocytes (5 x 105 cells per ml) for 48 hours in the presence of interleukin-2 and phytohaemagglutinin (P HA), followed by incubation on a 96-well multi-titre plates, and the mixture was incubated at 370C for a predetermined time (3 or 5 days).

(vi) The sample obtained in (v) was centrifuged for 10 minutes at 1000G, and the supernatant was filtered through a 0.45 um filter.

(vii) The filtrate obtained above was used for antigen measurement, and the residue was spread on a slide glass and subjected to testing by the fluorescent antibody technique.

(viii) As control samples (for sensitivity measurement), each agent was immersed in RPMI-1640 medium at concentrations of 10 1, 10 2, 10-3 10 and 10 5, each of the mixtures thus obtained was subjected to operations (ii) to (viii) above, and the samples thus prepared were submitted to testing by the fluorescent antibody technique.

(3) Verification of the effect to remove HIV and related compounds (A) Measurement of antigen concentration The concentration of antigen in culture liquid was determined according to the procedure given below using HIV antigen measuring kit (du Pont): (i) Test sample (180 ul) was mixed with 5% aqueous solution of Triton X100 (20 ul) on a microplate.

(ii) The mixture was allowed to stand overnight at room temperature.

(iii) Automatic washing with water was repeated three times.

(iv) A solution of biotin-treated antibody (100 v1) was added. and the mixture was incubated at 370C for two hours.

(v) Automatic washing.

(vi) HRP (horse-radish peroxidase)-sterptoavidin (100 ul) was added. and the mixture was incubated for 15 minutes.

(vii) After automatic washing, 100 ul of an aqueous solution of OPD (o-phenylenediamine) was added.

(viii) 4N-H2S04 solution (50 ul) was added to terminate the reaction.

(ix) Absorbence at 492 nm (620 nm for blank) was measured, and HIV concentration was calculated from a calibration curve.

The result obtained is summarized in Table 2.

As can be seen from the table, little of HIV antigen (P-24) was adsorbed by propylene, quartz and SO3 Na-type cation-exchange resin (substances having a neutral surface). On the other hand, the agents with a weakly acidic or weakly alkaline surface (particularly SO3H- and COOH-type cation-exchange resins and silica-alumina having a weakly acidic surface) adsorb HIV antigen efficiently.

Table 2

No. Agent (surface pH) Antigen Concn. (ng/ml) After 3 days After 5 days - Control T 0.520 0.942 Control ( - ) 0 0 Camp. Example 1 Polypropylene ( 7.0 ) 0.490 0.936 Gaw. Example 2 Quartz ( 7.0 ) 0.500 0.910 Comp. Example 3 IR-120B ( SO3Na-type ) 0.430 0.800 ( 7.0 ) Example 1 Hydroxyapatite ( 6.5 ) 0.351 0.570 Example 2 Hydroxyapatite ( 8.5 ) 0.189 0.294 Example 3 Silica-alunina ( 3.0 ) 0.074 0.122 Example 4 Anion-exchange resin 0.373 0.608 [ NH2-type ] ( 8.5 ) Example 5 Cation-exchange resin 0.114 0.164 [ COOH-type ] ( 3.0 ) Example 6 Cation-exchange resin 0 0 [ SO3H-type ] ( 3.0 ) Example 7 Alumina ( 5.0 ) 0.341 0.492 Example 8 Alumina ( 10.0 ) 0.180 0.272 (B) Evaluation by the fluorescent antibody technique Presence or absence of HIV and related compounds in the residue separated in (2) above (Procedure for removing HIV and related compounds) was determined by the fluorescent antibody technique as described below, and the effectiveness of the agents was evaluated based on the result obtained.

(i) Each of the residues obtained in the HIV adsorbing operation ((2)(i)-(vii)) was washed with 1 ml PBS for one minute at a low revolution speed.

(ii) The washed residue was suspended in 50 l PBS, and two spots of the suspension thus obtained were placed on a slide glass for fluorescent antibody measurement.

(iii) The spots on the slide glass were fixed by immersion in methanol.

(iv) The fixed spots were dehydrated by addition of acetone, followed by air drying.

(v) P-24 antibody - PBS solution (vol ratio; 1:40) (20 v1) was added to each spot.

(vi) The spots thus treated were incubated at 37 C for 20 minutes and then washed by PBS solution for 20 minutes.

(vii) Mouse anti-IgG antibody - PBS solution (vol ratio; 1:20) (20 1l1) was added to each spot.

(viii) The spots thus treated were incubated at 37 0C for 20 minutes and then washed by PBS solution for 20 minutes.

(ix) Streptoavidin/fluorescein - PBS solution (vol ratio; 1:50) (20 p1) was added to each spot.

(x) The spots thus treated were incubated at 37 0C for 10 minutes and then washed by PBS solution in the dark box.

(xi) One drop of a mixture of 90% glycerol and 10% PBS solution was added to each spot.

(xii) Presence or absence of fluorescent portions in each spot was observed with a microscope under ultraviolet irradiation.

The result is shown in Table 3.

Table 3

No. Agent ( surface pH ) Fluorescent antibody formation After 3 days @ After 5 days control ( + ) + + ------ Control ( - ) - Comp. Ex. 1 Polypropylene ( 7.0 ) + + Camp. Ex. 2 Quartz ( 7.0 ) + + Comp. Ex. 3 IR-120B ( SO3Na-type ) # + ( 7.0 ) Example 1 Hydroxyapatite ( 6.5 ) t + Example 2 Hydroxyapatite ( 8.5 ) f + Example 3 Silica-alumina ( 3.0 ) - Example 4 Anion-exchange resin # + [ NH2-type ] ( 8.5 ) Example 5 Cation-exchange resin - # [ COOH-type ] ( 3.0 ) Example 6 Cation-exchange resin - [ SO3H-type ] ( 3.0 ) Example 7 Alumina ( 5.0 ) - + Example 8 Alumina ( 10.0- ) + Diluted samples *:

C(+)x10-1: + C(+)x10-2:+ C(+)x10-3: + C(+)x10-4:+ C(+)x10-5:+ * Samples obtained by diluting Control (+) [ C(+) ] with RPMI medium to 10-1, 10-2, 10-3, 10-4 and 10-5 concentrations (after three days' incubation).

Antibody was detected in samples prepared by diluting Control (+) [C(+)] down to 10 5 concentration, indicating the high sensitivity of this method.

As can be seen from the table, anitbody production was negative (-) after three days' incubation in Examples 2, 3, 5, 6, 7 and 8. Though it was not negative in Examples 1 and 4, the production of antibody in these Examples was less than in the Comparative Examples. After five days' incubation, production of antibody was not positive on Examples 3, 5 and 6. On the other hand, production of antibody was observed on Examples 1, 2, 4, 7 and 8. This may be taken to indicate that an extremely small amount of HIV left unadsorbed produced a detectable quantity of antibody after five days' incubation, showing the slightly lower ability of these substances (Examples 1, 2, 4, 7 and 8) to remove HIV and related compounds than the others.

Further, evaluation of the ability to remove HIV and related compounds was carried out on Polypropylene (Comparative Example 1), silica-alumina (Example 3), cation-exchange resin (COOH-type) (Example 5) and cation-exchange resin (SO3 H-type) (Example 6). Culture supernatant of HIV-infected cells was treated with each of the agents in the same manner as stated in (2) except that the treating time of the culture supernatant with the agents was prolonged from 30 to 60 minutes in (2) (iii). Presence or absence of HIV and related compounds in the residues separated from the culture supernatants treated with the agents was determined after 7 days', 14 days' or 21 days' incubation by the immunofluorescence antibody measurement as described before. The result is shown in Table 4.

Table 4

Dilution ratio 10 10-1 10-2 10-3 10-4 10-5 Na Agent Days Comp Ex 1 Polypropylene 7 + + + + + + 14 + + + + + + 21 + + + + + + Example 3 Silica-alumina 7 - - I - /1 14 I I.

21 - - Example 5 Cation-exchange 7 - - resin(COOH- 14 II - I - type ) 21 - - Example 6 Cation-exchange 7 - - resin ( SO3H - 14 - - type ) 21 - - As can be seen from the table, the immunofluorescence antibody was not detected in Examples 3, 5 and 6. Therefore, it is indicated from this result that silica-alumina, cation-exchange resin (COOH-type) and cation-exchange resin (SO3H-type) have a high adsorption ability to HIV antigen.

(C) Evaluation of the ability to remove HIV and related compounds based on the intake of a thymidine derivative by juvenile lymphocytes The culture liquid, sensitized by CEM cells and treated above with the agents for removing HIV and related compounds, was tested for the intake of a thymidine derivative by juvenile lymphocytes according to the procedure given below.

(i) Lymphocytes taken from the cord blood of a new-born was diluted to a concentration of 5 x 105 cell/ml.

(ii) This suspension of juvenile lymphocytes was added to RPMI-164C medium containing 10% fresh bovine serum (FMS), 100 IU/ml penicillin, 100 ug/ml streptomycin, 1% PHA and 1 unitlml interleukin-2 (IL-2), and the mixture was incubated at 37 0C for 48 hours.

(ii) The culture liquid thus obtained (80 p1) was mixed with each of the samples sensitized with CEM cells and treated above with the agent for removing HIV and related compounds (20 ul), and the mixture was incubated for three or five days.

(iv) To the mixture incubated above, was added a radioactive thymidine derivative Imonoammonium (methyl-l ',2'-3H)-thymidine-5'-triphosphate, 3H-dTTP ] (1 uCuri/well) six hours before the end of incubation.

(v) The thus treated lymphocytes were collected on a Wattman glass filter by filtration, washed with 5% aqueous solution of trichloroacetic acid (10 ml) and then with methanol (10 ml), and air-dried.

(vi) The glass filter with lymphocytes collected thereon was put in a polyethylene container, an extraction agent (main component: toluene) was added. and the amount of 3H-dTTP taken in by the lymphocytes was measured by using a scintillation counter.

The ability of agents to remove HIV and related compounds was evaluated based on the intake of 3H-dTTP using the amount taken in by Control (-) [ C(-) ] as standard.

The result is shown in Table 5.

Table 5

No. Agent ( surface pH ) # Intake of H-dTTP (%) After 3 days After S days Control ( + ) 50.6 20.9 i Control ( - ) 100 100 Comp. Example 1 Polypropylene ( 7.0 ) 45.3 22.7 Comp. Example 2 Quartz ( 7.0 ) 46.0 22.0 Comp. Example 3 IR-120B ( SO3Na-type ) 43.0 25.0 ( 7.0 ) Example 1 Hydroxyapatite ( 6.5 ) 92.9 82.0 Example 2 Hydroxyapatite ( 8.5 ) 109.3 77.6 Fxmple 3 Silica-al ( 3.0 ) 96.1 78.4 Example 4 Anion-exchange resin 100.3 68.7 [ NH2-type ] ( 8.5 ) Example 5 Cation-exchange resin 103.2 91.9 [ COOH-type ] ( 3.0 ) Example 6 Cation-exchange resin 78.8 75.5 [ SO3H-type ] ( 3.0 ) Example 7 Alumina ( 5.0 ) 98.5 67.1 Example 8 | A1 ( 10.0 ) 98.5 72.3 As is apparent from Table 5, the intake of the thymidine derivative by Control (-) is about half that with Control (+) after three days of incubation. This indicates that lymphocytes are destroyed by HIV, leading to the reduction in the intake of thymidine derivative. This tendency is more marked after five days' incubation. When HIV culture liquid is treated with the agents for removing HIV and related compounds, on the other hand, no significant reduction in the intake of thymidine derivative is observed after three and five days' incubation, compared with Control (+) and neutral substances. This suggests that agents with a weakly acidic or alkaline surface have removed not only HIV but also those substances which participiate in the destruction of lymphocytes.

The result shout in Table 5 is nearly in agreement with the results of Tables 2 and 3. There are some agents, however, which are not so high in the ability of removing HIV (Table 2) but show high survival rate of lymphocytes in Table 5 (for example, Example 1). This also suggests the ability of agents to remove not only HIV but related compounds through adsorption.

(D) Evaluation of the ability to remove HIV antigen and related compounds by circulation system.

The HIV culture liquid containing flesh bovine serum (FBS) was circulated through a column filled with the above-mentioned agents, as stated below, and the HIV antigen concentration in the culture liquid was measured.

(i) 4 g of the agent was put in the column (volume 10 ml; inner diameter 15 mm) made of polypropylene and then the column was filled with saline solution.

(ii) The filled column was sterilized by autoclaving.

(iii) The saline solution in the sterilized column was replaced by RPMI-1640 medium adequately.

(iv) 50 ml of the HIV culture liquid (RPMI-1640 medium containing 10% FMS) (pH: 7.4) was circulated through the column at the rate of 7 ml/min for 1 hour.

(v) After the circulation the HIV antigen concentration in the culture liquid was measured.

The result is shown in Table 6.

Table 6

! IHIV antigen in Residual ratio ffa Agent the culture liquid of HIV antigen (ng/ml) (%) - Control (+) 80.00 100 Comp. Ex. 1 Polypropylene 76.96 96.2 Example 3 Silica-alumina 3.38 4.2 Example 5 Cation-exchange 3.14 3.5 resin(COOH-type) Example 6 Cation-exchange 2.78 3.9 resin(SO3H-type).

As can be seen from Table 6 the HIV antigen was eliminated effectively from the HIV culture liquid by the circulation system.

(4) Evaluation of safety of the agents.

Silica-alumina (Example 3), cation-exchange resin (COOH-type) (Example 5) and cation-exchange resin CSO3H-type) (Example 6) were treated in RPMI-1640 medium for 20 minutes at 121 0C to obtain an extract, respectively. Each of the extracts was added to the CEM cells and the cytotoxicity was examined. As a result, in all extracts, 90-95% of the CEM cells were viable after 21 days from the addition of the extract to the cells.

This indicates that the above agents are safe to living cells.

Comparative Example 4 Experiments similar to Example 1 were conducted using a cation-exchange resin Dowex 50WX8 (Dow Chemical; surface pH: 2.0) and an anion-exchange resin IRA-75 (Japan Organo Co; surface pH: 12.0). An increase in viscosity of the culture liquid was observed in both cases. This indicates that an undesirable phenomenon (coagulation of plasma) can occur when the plasma of patients is treated with these ion-exchange resins.

Hence, the use of substances with a strongly acidic or alkaline surface is not included in this invention.

As is apparent from the foregoing, HIV and related compounds can be efficiently removed by the method of this invention with no adverse effect upon the growth of normal lymphocytes. Use of the agents of this invention by the intracorporeal blood circulation system removes HIV and related compounds from the body fluid of patients in the aggravating period (in which a rapid increase in the amount of HIV is observed), thereby preventing the worsening of patients' conditions and so prolonging their lives. This method may also serve as an auxiliary means to medicinal therapies (for example, administration of antiviral agents). It is also possible to use the agents for purification of blood preparations and for concentration of HIV from blood samples for clinical tests.

Claims (6)

1. A solid substance with a weakly acidic or weakly alkaline surface for use in the manufacture of agents for removing human immunodeficiency virus and its related compounds contained in body fluid.

2. A solid cation-exchange resin of COOH- or SO3H-type for use in the manufacture of agents for removing human immunodeficiency virus and its related compounds contained in body fluid.

3. A solid silica-alumina for use in the manufacture of agents for removing human immunodeficiency virus and its related compounds contained in body fluid.

4. A method of manufacturing agents for removing human immunodeficiency virus and its related compounds contained in body fluid, in which a solid substance with a weakly acidic or weakly alkaline surface is used.

5. A method of manufacturing agents for removing human immunodeficiency virus and its related compounds contained in body fluid, in which a cation-exchange resin of COOH- or SO 3H -type is used.

6. A method of manufacturing agents for removing human immunodeficiency virus and its related compounds contained in body fluid, in which silica-alumina is used.