Engineering an improved adenovirus packaging cell line

Recombinant human adenovirus (Ad) vectors are being extensively explored for their use in gene therapy and recombinant vaccines. Ad vectors are attractive for many reasons, including the fact that (1) they are relatively safe, based on their use as live oral vaccines, (2) they can accept large transgene inserts, (3) they can infect dividing and postmitotic cells, and (4) they can be produced to high titers. However, there are also a number of major problems associated with Ad vectors, including transient foreign gene expression due to host cellular immune responses, problems with humoral immunity, and the creation of replication competent adenoviruses (RCA). Most Ad vectors contain deletions in the E1 region that allow for insertion of a transgene. However, the E1 gene products are required for replication and thus must be supplied in trans by a helper ceillille that will allow for the growth and packaging of the defective virus. For this purpose the 293 cell line (Graham et al., 1977) is used most often; however, homologous recombination between the vector and the cell line often results in the generation of RCA. The presence of RCA in batches of adenoviral vectors for clinical use is a safety risk because tlley . may result in the mobilization and spread of the replication-defective vector viruses, and in significant tissue damage and pathogenicity. The present research focused on the alteration of the 293 cell line such that RCA formation can be eliminated. The strategy to modify the 293 cells involved the removal of the first 380 bp of the adenovirus genome through the process of homologous recombination. The first step towards this goal involved identifying and cloning the left-end cellular-viral jUl1ction from 293 cells to assemble sequences required for homologous recombination. Polymerase chain reaction (PCR) was performed to clone the junction, and the clone was verified through sequencing. The plasn1id PAM2 was then constructed, which served as the targeting cassette used to modify the 293 cells. The cassette consisted of (1) the cellular-viral junction as the left-end region of homology, (2) the neo gene to use for positive selection upon tranfection into 293 cells, (3) the adenoviral genome from bp 380 to bp 3438 as the right-end region of homology, and (4) the HSV-tk gene to use for negative selection. The plasmid PAM2 was linearized to produce a double strand break outside the region of homology, and transfected into 293 cells using the calcium-phosphate technique. Cells were first selected for their resistance to the drug G418, and subsequently for their resistance to the drug Gancyclovir (GANC). From 17 transfections, 100 pools of G418f and GANCf cells were picked using cloning lings and expanded for screening. Genomic DNA was isolated from the pools and screened for the presence of the 380 bps using PCR. Ten of the most promising pools were diluted to single cells and expanded in order to isolate homogeneous cell lines. From these, an additional 100 G41Sf and GANef foci were screened. These preliminary screening results appear promising for the detection of the desired cell line. Future work would include further cloning and purification of the promising cell lines that have potentially undergone homologous recombination, in order to isolate a homogeneous cell line of interest.

Anne McPherson family fonds, 1852-2011 [non-inclusive]

Frank C. (Case) McCordick (1873-1946) was the son of William Henry (1849-1930) and Emily D. Howell (1851-1927) McCordick. William H. McCordick was in the coal business. The McCordick family included Frank Case, Mabel Gertrude, Ethel Howell and Arthur Stanley. Frank C. McCordick was educated in St. Catharines, and worked with his father in the coal business and eventually opened up a leather tanning operation. McCordick was active in the Lincoln Regiment and in 1906 was promoted to captain and in command of Company A, 19th Regiment. He was promoted to major and at the outbreak of war he was sent overseas as a commander of the 35th Battalion of the Canadian Expeditionary Forces (CEF). Upon arrival in France he was made officer commanding the 15th Battalion, King’s Own Yorkshire Light Infantry (KOYLI). After the war and his return to Canada he continued to play an active role in the local military units in the area as well as in Hamilton. After his retirement from the military in 1927 McCordick served as alderman and then mayor of St. Catharines from 1930 to 1931. He was a member of a large number of civic clubs, including St. Catharines Chamber of Commerce, Y.M.C.A., Lion’s Club, St. Catharines Golf Club, Detroit Boat Club, the St Catharines Club, as well as a member of several Masonic lodges. He continued to operate McCordick Tannery and other local investments. In 1903 Frank C. McCordick married May Beatrice Simson, daughter of Thomas E. Simson of Thorold. They had three children, E. (Edward) Frank McCordick, Bruce McCordick and (Margaret) Doris McCordick (m. Hubert Grigaut, d. 1977). The McCordick family resided at 82 Yates Street, near Adams Street. May Simson McCordick (b. 1873) was the daughter of Thomas Edward (1836-1908) and Julia Headlam (1844-1887) Simson of Thorold. Her siblings included: Edward, Frances, John, Augusta, Georgia and Gertrude. E. (Edward) Frank McCordick (1904-1980) was born in St. Catharines, Ont., attended Lake Lodge School in Grimsby, Ridley College in St. Catharines, Beechmont Preparatory School in England, Upper Canada College in Toronto and graduated from Royal Military College in Kingston, Ont. in 1925. Upon graduation he was made a lieutenant in the 10th (St. Catharines) Field Battery. In 1929 he married Helen Stanley Smith, daughter of Stanley George and Mary Walker Smith of St. Catharines. Col. McCordick, now promoted to Major, played an active role in the 10th (St. Catharines) Field Battery, being officer commanding the battery. In late 1939 McCordick headed to England for artillery tactical training and on December 6, 1939 the battery began the long trek overseas. McCordick saw action in Italy and in Holland. Upon his return to Canada at the end of the war he was the Liberal candidate in the federal election for Lincoln County. He remained active in the local military serving as honorary lieutenant-colonel of the 56th Field Regiment (ARCA) and in 1976 as the honorary colonel of the regiment. Col. McCordick held the Efficiency Decoration, the Order of the British Empire, granted in 1945 and was made an officer in the Order of St. John in 1978. He continued to serve his community in various capacities, including the Unemployment Insurance Canada Board, Royal Trust Company and the St. John Ambulance Society. He remained an active member of the alumni of Royal Military College, editing and compiling a newsletter and organizing reunion weekends. He kept in close contact with many of his classmates. Helen Stanley Smith McCordick lived in St. Catharines, Ont., attended Robertson School, and graduated from the University of Toronto in 1926 with a Bachelor of Arts degree in Modern Languages. During the war years (1939-1945) Helen was active in the Transport division of the local branch of the Canadian Red Cross and the Women’s Auxiliary of the 10th Field Battery. In 1932 E. Frank and Helen McCordick welcomed their only child, (Catharine) Anne McCordick. Helen continued to play an active role in her community until her passing in 1997. Stanley George Smith (1865-1960) was born in St. Catharines, Ont., the only child of William Smith (d. June 16, 1876) a native of Edinburgh, Scotland and his wife Hannah Louisa Maria Bulkeley a native of Fairfield, Connecticut. Stanley George Smith married Mary Walker of Guelph, Ont.(d. 1956) Mary was the daughter of Hugh and Elizabeth (d. 1924) Walker. Her siblings included Margaret, Agnes, Jessie, Isabella, Lorne, Ada, Alice, Eva, Alexander and George. Hugh Walker was a prominent fruit and vegetable merchant in Guelph. On 1904 their only child, Helen Stanley Smith was born. He was a post office clerk, and the treasurer for the James D. Tait Co. Ltd., a clothing and dry goods retailer in St. Catharines. The family lived at 39 Church Street in St. Catharines, Ont.

Niagara Falls Park: Notice to Treat to Sutherland Macklem, Esquire, 1885

The Niagara Parks Commission administrative headquarters are located in Oak Hall which is on the cliff above Dufferin Islands. In 1798 this land was granted by the crown to United Empire Loyalist James Skinner until 1898. A century later it was the home of the Clarks, Streets and Macklems. These families controlled the mills of Bridgewater which was a pioneer industrial village located at Dufferin Islands. Then, it was known as Clark Hill. Colonel Thomas Clark, commander of the Second Lincoln Militia in the War of 1812 is the earliest known occupant of the house. When Clark died in 1837, the house went to Thomas Clark Street who was the son of the Colonel’s partner. Mr. Street was a bachelor and his sister, widow of Dr. T.C. Macklem, managed his household. Mrs. Macklem had 2 sons. The eldest son drowned in the Niagara River at the age of 8 and the younger son, Sutherland became heir to the estate. Mr. Macklem opened Cynthia Islands and Cedar Island to the public and had roads built to reach them. Two suspension bridges connected them to the mainland and tolls were charged on the bridges. The improvements to the land cost Macklem $18,962. He called the bridges “Bridge Castor” and “Bridge Pollux”. There was also an office built at the end of Bridge Castor. Macklem also spent $454 fixing up the Burning Spring Building (the burning spring is enclosed in a barrel which collects gas and lets it out through a tube at the top). Macklem received a yearly income of $56,378.79 from tourists and visitors. In 1887 Cynthia Islands and Cedar Island were deeded to the crown and became part of Queen Victoria Park. The name Cynthia was changed to Dufferin in honour of Lord Dufferin. Sources: www.niagarafrontier.com/parks.html www.niagarafrontier.com/burningsprings.html

Indenture between Charles Adams and Charles Ira Ames

Indenture of mortgage between Charles Adams of Hamilton and Charles Ira Ames of Hamilton of the first part to Frances Malvinia, wife of Charles Adams of the second part and Thomas Clarke Street of Chippewa, eldest son and heir of Samuel Street of the third part regarding Lot no. 5 in the 5th Concession in the Township of Caradoc in the County of Middlesex, May 5, 1854.